CN111713410A - Kiwi explant detoxification method - Google Patents
Kiwi explant detoxification method Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
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- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
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Abstract
The invention provides a kiwifruit explant detoxification method, which sequentially comprises the following steps: selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, cleaning, sterilizing, sucking dry with sterile filter paper, and stripping meristematic tissues of explants; and (3) induction culture: inoculating the meristem of the explant to an induction culture medium, and performing dark culture and light dark culture to obtain cluster buds; rooting culture: when the cluster buds grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark and light to obtain tissue culture seedlings; hardening and transplanting seedlings: culturing tissue culture seedling, transplanting to mixed matrix, and culturing to obtain detoxified seedling. According to the invention, strong virus-free seedlings are obtained through explant selection, induction culture, rooting culture and seedling hardening and transplanting in sequence, the production cost is reduced, the seedling quality is improved, the problems of virus carrying and high virus carrying rate of kiwi seedlings in the prior art are effectively solved, and the large-area popularization of the kiwi planting industry is facilitated.
Description
Technical Field
The invention belongs to the technical field of tissue culture, and particularly relates to a method for detoxifying a kiwi fruit explant.
Background
Kiwi fruit, a plant of the genus Actinidia of the family Actinidiaceae, is native to Chinese vine fruit trees and is an important economic fruit tree. The kiwi fruit is rich in various vitamins, especially high in vitamin C, and also contains a large amount of minerals, arginine and the like, so the kiwi fruit is also called as the king of fruit. The kiwi fruit is widely applied to food and beverage processing, health products and other forms. The virus disease of the kiwi fruit, which is a perennial fruit tree, is a very serious disease, no effective treatment measure and method exist at present, and the method has great resistance to popularization of kiwi fruit planting.
The seed propagation of the kiwi fruit is easy to cause variety degeneration, some excellent characters of the female parent are difficult to maintain, and the kiwi fruit is easy to carry pathogens such as virus diseases, canker diseases and the like through cuttage or grafting. The seedlings obtained by the conventional method are often toxic, the survival rate is low, strong kiwi seedlings cannot be obtained, the planting cost is high, and the seedling quality is low.
Disclosure of Invention
Aiming at the problems in the prior art, the invention provides a kiwi fruit explant detoxification method, which comprises explant selection, induction culture, rooting culture and seedling hardening transplantation in sequence to obtain robust detoxified seedlings, so that the production cost is reduced, the seedling quality is improved, the problems of high toxicity carrying rate and the like of kiwi fruit seedlings in the prior art are effectively solved, and the large-area popularization of the kiwi fruit planting industry is facilitated.
In order to achieve the purpose, the technical scheme adopted by the invention for solving the technical problems is as follows: the method for detoxifying the kiwi fruit explant sequentially comprises the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, cleaning, sterilizing, sucking with sterile filter paper, and stripping 2-2.5 mm explant meristem;
(2) and (3) induction culture: inoculating the meristem of the explant obtained in the step (1) to an induction culture medium, performing dark culture for 1-2 days, and performing light-dark culture for 25-35 days to obtain cluster buds;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings;
(4) hardening and transplanting seedlings: and (4) culturing the tissue culture seedlings obtained in the step (3) for 2-4 days at 23-28 ℃ and 60-65% humidity, removing the culture medium on the tissue culture seedlings, transplanting the tissue culture seedlings into a mixed matrix, and culturing to obtain the virus-free seedlings.
Further, the explant is a shoot tip or axillary bud.
Further, in the step (1), during cleaning and disinfection, the water is firstly washed for 1-2 hours, then the water is soaked in 75 vt% ethanol for 20-40 s, the water is washed for 2-4 times by using sterile water, then the water is disinfected for 2-4 min by using 3-5 wt% sodium hypochlorite solution, the water is washed for 5-7 times by using the sterile water, and finally the water is sucked and dried by using sterile filter paper.
Further, in the light-dark culture in the steps (2) to (3), the culture is carried out for 12 hours under 2000Lx illumination and 23-26 ℃ every day, and the culture is carried out for 12 hours under 21-23 ℃ in the dark.
Further, 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar are added into the induction culture medium on the basis of 1L MS culture medium; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L.
Further, the rooting culture medium is added with naphthylacetic acid, sucrose and agar on the basis of 1L1/2MS culture medium; wherein the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L.
Further, the mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 1-2: 1:1 to obtain a mixed matrix.
Further, when the seedlings are hardened in the step (4), the illumination is 20000 to 21000 Lx.
In summary, the invention has the following advantages:
1. the detoxification tissue culture of herbaceous plants can select shorter and smaller stem tips to ensure the detoxification effect, but in woody plants rich in phenolic substances, kiwi fruit is in the front, so that tissue culture seedlings are easy to brown, and dead seedlings are easy to cause. According to the invention, the meristem of 2-2.5 mm stem tip or axillary bud is stripped, the length is easy to survive, and the virus removal effect can be considered.
2. In addition to the stem tip, the axillary buds are innovatively used as material taking objects, and are easier to separate than the stem tip, so that the operation is simplified, the number of the axillary buds is large, more material sources are provided, and the method is favorable for large-scale production and application. We find that axillary buds and stem tips have the same effect in the detoxification effect and the callus culture process, and the quality of the detoxified seedlings can be ensured.
3. Compared with other woody plants, the kiwi fruit has low explant activity, is very easy to brown under the action of phenolic substances and external environment, and has extremely high requirements on a culture medium formula. Sodium hypochlorite has certain destructive effect on explants, and experiments show that the browning can be effectively inhibited by controlling the concentration of the sodium hypochlorite to be less than 4% during the disinfection by using the sodium hypochlorite. The duration of the sterilization also has an effect on browning, but not as much as the concentration of the sterilizing agent. The disinfection effect of soaking the material with 1.5% sodium hypochlorite for 45min is similar to that of 3% sodium hypochlorite treatment for 3min in the invention, and the invention provides the best disinfection method for the kiwi explant.
4. The concentration ratio of 6BA and NAA in various culture media involved in obtaining the kiwifruit virus-free seedlings is also greatly different from that of herbaceous plants, and the research finds that the relative ratio of auxin/cytokinin is smaller and is favorable for adventitious bud proliferation, and the relative ratio of auxin/cytokinin is larger and is favorable for rooting. The rooting effect of the kiwi fruit is better when the relative dosage of the NAA is adjusted to an extremely low concentration (NAA concentration of 0.2 mg/L).
5. According to the research of the invention, after the callus is inoculated on the culture medium, dark culture is carried out for 1-2 days, and then illumination culture is carried out, so that the growth speed of the explant is properly delayed in the initial stage, the rapid accumulation of phenolic substances can be effectively reduced, the phenolic substances are inhibited from being converted into brown quinone substances, and the browning of kiwi fruit tissues is inhibited. In addition, the light and the dark are alternately 12h respectively, and the illumination of 2000lx needs to be accurately controlled.
6. The degree of the age and the young of the kiwi fruit explants also determines the difficulty and the later pollution condition of disinfection. The research of the invention finds that the aged explant has obviously higher pollution rate than the young explant. The old kiwi fruit leaves and axillary buds are lignified by dense villi and the old kiwi fruit stems, so that the old kiwi fruit has more bacteria and is difficult to disinfect. The present invention suggests the isolation of young, tender, non-lignified kiwifruit as the primary source of material.
Drawings
FIG. 1 is a schematic representation of dissected microscopic shoot tips or axillary buds meristems upon dissection;
FIG. 2 is a schematic diagram showing the induced differentiation of shoot tip or axillary bud meristem into small buds during induction culture;
FIG. 3 is a schematic view in rooting culture;
FIG. 4 is a schematic of a detoxified shoot.
Detailed Description
Example 1
A kiwi explant detoxification method sequentially comprises the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, washing with running water for 1h, soaking for 20s with 75 vt% ethanol, washing with sterile water for 2 times, disinfecting with 3 wt% sodium hypochlorite solution for 2min, washing with sterile water for 5 times, sucking with sterile filter paper, and peeling 2-2.5 mm stem tips or axillary bud meristems;
(2) and (3) induction culture: inoculating the stem tip or axillary bud meristem obtained in the step (1) to an induction culture medium, culturing for 1d in dark, and culturing for 25d in light and dark to obtain cluster buds; culturing in dark at 2000Lx light and 23 deg.C for 12 hr, and culturing in dark at 21 deg.C for 12 hr; the induction culture medium is prepared by adding 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L MS culture medium; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings; culturing in dark at 2000Lx light and 23 deg.C for 12 hr, and culturing in dark at 21 deg.C for 12 hr; the rooting culture medium is added with naphthylacetic acid, sucrose and agar on the basis of 1L1/2MS culture medium; wherein the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L;
(4) hardening and transplanting seedlings: culturing the tissue culture seedlings obtained in the step (3) for 2d under the conditions of 20000Lx, 23 ℃ and 60% humidity, removing the culture medium on the tissue culture seedlings, transplanting the tissue culture seedlings into a mixed matrix, and culturing to obtain detoxified seedlings; the mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 2:1:1 to obtain the mixed matrix.
Example 2
A kiwi explant detoxification method sequentially comprises the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, washing with running water for 1h, soaking for 30s with 75 vt% ethanol, washing with sterile water for 3 times, disinfecting with 4 wt% sodium hypochlorite solution for 3min, washing with sterile water for 6 times, sucking with sterile filter paper, and peeling 2-2.5 mm stem tips or axillary bud meristems;
(2) and (3) induction culture: inoculating the stem tip or axillary bud meristem obtained in the step (1) to an induction culture medium, culturing for 1d in dark, and culturing for 30d in light and dark to obtain cluster buds; culturing in dark at 2000Lx light and 24 deg.C for 12 hr, and culturing in dark at 22 deg.C for 12 hr; the induction culture medium is prepared by adding 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L MS culture medium; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings; culturing in dark at 2000Lx light and 24 deg.C for 12 hr, and culturing in dark at 22 deg.C for 12 hr; rooting culture medium is prepared by adding Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L1/2MS culture medium; wherein the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L;
(4) hardening and transplanting seedlings: culturing the tissue culture seedlings obtained in the step (3) for 3d under the conditions of 20000Lx, 24 ℃ and 62% humidity, removing the culture medium on the tissue culture seedlings, transplanting the tissue culture seedlings into a mixed matrix, and culturing to obtain detoxified seedlings; the mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 2:1:1 to obtain the mixed matrix.
Example 3
A kiwi explant detoxification method sequentially comprises the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, washing with running water for 1h, soaking for 35s with 75 vt% ethanol, washing with sterile water for 3 times, disinfecting with 4 wt% sodium hypochlorite solution for 3min, washing with sterile water for 6 times, sucking with sterile filter paper, and peeling 2-2.5 mm stem tips or axillary bud meristems;
(2) and (3) induction culture: inoculating the stem tip or axillary bud meristem obtained in the step (1) to an induction culture medium, culturing in dark for 1d, and culturing in light and dark for 32d to obtain cluster buds; culturing in dark at 2000Lx light and 25 deg.C for 12 hr, and culturing in dark at 22 deg.C for 12 hr; the induction culture medium is prepared by adding 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L MS culture medium; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings; culturing in dark at 2000Lx light and 25 deg.C for 12 hr, and culturing in dark at 22 deg.C for 12 hr; rooting culture medium is prepared by adding Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L1/2MS culture medium; wherein the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L;
(4) hardening and transplanting seedlings: culturing the tissue culture seedlings obtained in the step (3) for 3d under the conditions of 20000Lx, 26 ℃ and 64% humidity, removing the culture medium on the tissue culture seedlings, transplanting the tissue culture seedlings into a mixed matrix, and culturing to obtain detoxified seedlings; the mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 2:1:1 to obtain the mixed matrix.
Example 4
A kiwi explant detoxification method sequentially comprises the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, washing with running water for 2h, soaking with 75 vt% ethanol for 40s, washing with sterile water for 4 times, disinfecting with 5 wt% sodium hypochlorite solution for 4min, washing with sterile water for 7 times, sucking with sterile filter paper, and peeling 2-2.5 mm stem tips or axillary bud meristems;
(2) and (3) induction culture: inoculating the stem tip or axillary bud meristem obtained in the step (1) to an induction culture medium, culturing in dark for 2d, and culturing in light and dark for 35d to obtain cluster buds; culturing in dark at 2000Lx light and 26 deg.C for 12 hr, and culturing in dark at 23 deg.C for 12 hr; the induction culture medium is prepared by adding 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar based on 1L MS culture medium; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings; culturing in dark at 2000Lx light and 26 deg.C for 12 hr, and culturing in dark at 23 deg.C for 12 hr; the rooting culture medium is added with naphthylacetic acid, sucrose and agar on the basis of 1L1/2MS culture medium; wherein the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L;
(4) hardening and transplanting seedlings: culturing the tissue culture seedling obtained in the step (3) for 4 days under the conditions of 21000Lx, 28 ℃ and 65% humidity, removing the culture medium on the tissue culture seedling, transplanting the tissue culture seedling into a mixed matrix, and culturing to obtain a detoxified seedling; the mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 2:1:1 to obtain the mixed matrix.
Experimental example 1
1mm, 2mm, 2.5mm, 3mm and 4mm shoot tips or axillary bud meristems are respectively selected, cultured according to the detoxification method of the embodiment 3, each group is subjected to 3 parallel experiments, and finally the pollution rate, the induction rate and the detoxification rate are measured, and the results are shown in table 1.
TABLE 1 contamination rate, induction rate and detoxification rate of meristems of shoot tips or axillary buds of different sizes
| Size of stem tip or axillary bud/mm | Percent of contamination/%) | Induction rate/% | Detoxification rate/% |
| 1 | 0 | 10 | 91 |
| 2 | 4 | 76 | 75 |
| 2.5 | 16 | 85 | 70 |
| 3 | 19 | 87 | 46 |
| 4 | 20 | 91 | 30 |
As can be seen from Table 1, under the same treatment method and environment, the kiwi seedlings cultured by 1mm stem tips have the highest detoxification rate, but have the lowest induction rate, so that the kiwi seedlings are not adopted. The selection of 3mm or more stem tips has a considerable induction rate, but the detoxification rate is below 50%, which is contrary to the original purpose of the experiment. Therefore, the size of the optimal stem tip explant is 2-2.5 mm, and the good detoxification effect can be ensured while most explants are normally differentiated.
Experimental example 2
Carrying out culture experiments according to the detoxification method in the example 3, selecting different induction culture media for each group, and carrying out 3 groups of parallel experiments for each group; the MS culture medium, the sucrose and the agar in the induction culture medium have the same addition amount, and the preparation process is the same; wherein, the contents of 6-benzylaminopurine and Naphthalene Acetic Acid (NAA) in each group are shown in Table 2. The number of adventitious buds and the induction rate were determined after completion of the culture, and the results are shown in Table 2.
TABLE 2 adventitious bud number and Induction Rate for different Induction media
As can be seen from Table 2, in the same treatment method, the formulation of the MS culture medium, 6-BA 2.0mg/L and NAA 0.1mg/L culture medium induces the most adventitious buds, ensures that more explant materials can be obtained during rooting culture, and is the most ideal culture medium for inducing proliferation.
Experimental example 3
The culture experiment is carried out according to the detoxification method of the embodiment 3, each group selects different rooting culture media, and each group is carried out with 3 groups of parallel experiments; the MS culture medium, the sucrose and the agar in the rooting culture medium have the same addition amount, and the preparation process is the same; wherein the content of each group of Naphthalene Acetic Acid (NAA) is shown in Table 3. The number of adventitious buds and the induction rate were determined after completion of the culture, and the results are shown in Table 3.
TABLE 3 rooting percentage and transplanting survival rate of different rooting media
| Treatment of | NAA/mg/L | Rooting percentage/%) | Survival rate of transplantation/%) |
| 1 | 0 | 10 | 30 |
| 2 | 0.1 | 80 | 85 |
| 3 | 0.2 | 86.7 | 90 |
| 4 | 0.4 | 50 | 60 |
As can be seen from Table 3, under the same treatment method, the root system induced by the 1/2MS + NAA 0.2mg/L culture medium formula is more developed, which ensures that the tissue culture seedling can grow better after being transplanted, and is the most ideal rooting culture medium.
While the present invention has been described in detail with reference to the illustrated embodiments, it should not be construed as limited to the scope of the present patent. Various modifications and changes may be made by those skilled in the art without inventive step within the scope of the appended claims.
Claims (8)
1. A kiwi explant detoxification method is characterized by sequentially comprising the following steps:
(1) selecting an explant: removing leaves of kiwi fruit seedlings, cutting 1-2 cm stem segments with buds, cleaning, sterilizing, sucking with sterile filter paper, and stripping 2-2.5 mm explant meristem;
(2) and (3) induction culture: inoculating the meristem of the explant obtained in the step (1) to an induction culture medium, performing dark culture for 1-2 days, and performing light-dark culture for 25-35 days to obtain cluster buds;
(3) rooting culture: when the cluster buds obtained in the step (2) grow to 1-2.5 cm and are accompanied with 2-5 new leaves, transferring the cluster buds to a rooting culture medium, and culturing in the dark for 30 days to obtain tissue culture seedlings;
(4) hardening and transplanting seedlings: and (4) culturing the tissue culture seedlings obtained in the step (3) for 2-4 days at 23-28 ℃ and 60-65% humidity, removing the culture medium on the tissue culture seedlings, transplanting the tissue culture seedlings into a mixed matrix, and culturing to obtain the virus-free seedlings.
2. The method for detoxification of kiwi explant of claim 1, wherein said explant is shoot tip or axillary bud.
3. The method for detoxifying the kiwi fruit explant according to claim 1, wherein in the step (1), the kiwi fruit explant is washed with running water for 1-2 hours, then soaked in 75 vt% ethanol for 20-40 seconds, washed with sterile water for 2-4 times, then disinfected with 3-5 wt% sodium hypochlorite solution for 2-4 min, washed with sterile water for 5-7 times, and finally dried by sterile filter paper.
4. The method for detoxifying an explant of kiwi fruit according to claim 1, wherein in the culturing in darkness in steps (2) - (3), the explant is cultured in 2000Lx light at 23-26 ℃ for 12h every day, and is cultured in darkness at 21-23 ℃ for 12 h.
5. The method for detoxification of kiwi explants of claim 1, wherein said induction medium is based on 1L MS medium, added with 6-benzylamino adenine, Naphthalene Acetic Acid (NAA), sucrose and agar; wherein, the concentration of 6-benzylamino adenine is 2mg/L, the concentration of naphthylacetic acid is 0.2mg/L, the concentration of sucrose is 30g/L, and the concentration of agar is 7 g/L.
6. The method for detoxifying a kiwi explant according to claim 1, wherein said rooting medium is based on 1L1/2MS medium, and Naphthalene Acetic Acid (NAA), sucrose and agar are added; wherein, the concentration of the naphthylacetic acid is 0.2mg/L, the concentration of the sucrose is 15g/L, and the concentration of the agar is 7 g/L.
7. The method for detoxification of kiwi explant of claim 1, wherein said mixed matrix is prepared by the following method: mixing turfy soil, vermiculite and perlite according to the volume ratio of 1-2: 1:1 to obtain a mixed matrix.
8. The method for detoxifying a kiwi fruit explant according to claim 1, wherein during hardening off in step (4), illumination is 20000 to 21000 Lx.
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| CN114051882A (en) * | 2021-10-22 | 2022-02-18 | 湖北省农业科学院果树茶叶研究所 | Production method of kiwi fruit seedlings without canker pathogenic bacteria |
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| CN114051882A (en) * | 2021-10-22 | 2022-02-18 | 湖北省农业科学院果树茶叶研究所 | Production method of kiwi fruit seedlings without canker pathogenic bacteria |
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