CN110301278A - One kind " Red Male " Kiwi berry is without Pathogenic Fungus of Canker seedling fostering technology - Google Patents
One kind " Red Male " Kiwi berry is without Pathogenic Fungus of Canker seedling fostering technology Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01G17/00—Cultivation of hops, vines, fruit trees, or like trees
- A01G17/005—Cultivation methods
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
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- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
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Abstract
The invention belongs to fruit rearing new variety technical fields, especially a kind of " Red Male " Kiwi berry is without Pathogenic Fungus of Canker seedling fostering technology, operation for current prevention and treatment canker mode needs to be grasped certain experiences, and compare labor intensive, energy problem, it is proposed that following scheme includes the following steps, the Fiber differentiation of " Red Male " Kiwi berry bud, it takes Chinese gooseberry garden to take branch or blade, by surface sterilization, budding is induced on culture medium；The proliferation of " Red Male " Kiwi berry bud expands the quantity of bud to the obtained bud of induction into proliferation；The removing of Prospect on Kiwifruit Bacterial Canker pathogen under anatomical lens, separates shoot tip meristem, is cultivated to obtain sprouting, and each bud is numbered and is proliferated using the bud of proliferation as material.The present invention cuts off the propagation of pathogen from source, can effectively prevent the generation of Prospect on Kiwifruit Bacterial Canker, sprays insecticide compared to rising, and more environmentally-friendly health compares grafting plant, and convenient operation and management is conducive to cultivate.
Description
Technical field
The present invention relates to fruit rearing new variety technical fields, more particularly to a kind of " Red Male " Kiwi berry is without canker cause of disease
Strain seedling Cultivating techniques.
Background technique
Kiwi berry is one of important fruit in the world, is rich in various trace elements, has multiple efficacies, wherein " Red Male "
Red cyanidin in Kiwi berry pulp also has anti-oxidant, anti-aging, pre- anti-cancer, enhancing eyesight and improvement sleep etc. a variety of
Effect, it is deep to be liked by people all over the world.
Prospect on Kiwifruit Bacterial Canker is a kind of crushing bacteriosis for endangering Kiwi berry, seriously affects the growth of Kiwi berry, is produced
Amount and quality, or even cause plant dead, and " Red Male " Kiwi berry is more sensitive to canker, is easy morbidity, prevention and treatment is burst at present
It is the absorption rate improved by improvement soil texture tree body to multiple element, limitation fruiting amount that ulcer disease, which uses most modes,
To reduce injured probability and to pocket knife, in lesions position scarfing brush medicament, operation needs to be grasped certain experiences, and compares
Labor intensive, energy, we have proposed a kind of " Red Male " Kiwi berrys without Pathogenic Fungus of Canker seedling fostering technology thus.
Summary of the invention
Operation based on current prevention and treatment canker mode needs to be grasped certain experiences, and compares labor intensive, energy technology
Problem, the invention proposes a kind of " Red Male " Kiwi berrys without Pathogenic Fungus of Canker seedling fostering technology.
One kind " Red Male " Kiwi berry proposed by the present invention includes the following steps: without Pathogenic Fungus of Canker seedling fostering technology
S1: the Fiber differentiation of " Red Male " Kiwi berry bud takes Chinese gooseberry garden to take branch or blade, by surface sterilization, is training
It supports and induces budding on base；
S2: the proliferation of " Red Male " Kiwi berry bud expands the quantity of bud to the obtained bud of induction into proliferation；
S3: the removing of Prospect on Kiwifruit Bacterial Canker pathogen under anatomical lens, separates shoot tip meristem using the bud of proliferation as material,
It is cultivated to obtain sprouting, and each bud is numbered and is proliferated；
S4: the screening of the bud without Prospect on Kiwifruit Bacterial Canker pathogen, Bacteria Culture identification, by the fertile material of the obtained each bud of S3
It carrying out grinding to be slurried, is coated on bacteria culture media, is cultivated in biochemical cultivation case, observation has abacterial growth, and
It is control with Prospect on Kiwifruit Bacterial Canker pathogen, is compared, filters out the bud of no bacterial growth；PCR identification, with no bacterial growth
Bud extract DNA, and as pcr template, carry out PCR reaction with the special primer of Prospect on Kiwifruit Bacterial Canker pathogen DNA, then right
PCR product is separated by electrophoresis, and whether observation has the specific product of Prospect on Kiwifruit Bacterial Canker pathogen, filters out no specific product
Bud；
S5: expanding the bud of numerous no Prospect on Kiwifruit Bacterial Canker pathogen, to S4 by Bacteria Culture and PCR method screen without macaque
The bud of peach Pathogenic Fungus of Canker is proliferated, and a large amount of bud is obtained；
S6: S5 is expanded numerous obtained bud and is inoculated on root media and carries out by the regeneration without Prospect on Kiwifruit Bacterial Canker pathogen plant
It takes root, to obtain completely without Prospect on Kiwifruit Bacterial Canker pathogen plant；
S7: the regenerated plant of S6 is carried out adaptability by the domestication without Prospect on Kiwifruit Bacterial Canker pathogen plant in being transplanted to greenhouse
Training after one month, moves on to outdoor carry out nature cultivation, forms commercial seedling.
Preferably, the culture medium includes MS culture medium, 1/2MS culture medium, WPM culture medium and B5 medium, the training
Feeding base sterilizes 15 min, every liter of 7g containing agar, sucrose 20g at 121 DEG C；Adjust pH value between 5.8-6.0, wherein 1/2MS
Culture medium configuration is to halve a great number of elements content for configuring MS culture medium, and every liter of addition sucrose 20g, other contents are identical, energy
Enough hestening rootings.
Preferably, in the S1, surface sterilization is cleaned once under the tap water that flows slowly with detergent, is rinsed once, sufficiently
It is transferred in superclean bench after cleaning, respectively in the liquor natrii hypochloritis containing 1% effective chlorine and containing 20 Hes of disinfection in 0.2% polysorbas20
Then 15 min are used rinsed with sterile water 3 times.
Preferably, the condition of the culture are as follows: cultivation temperature is 1 DEG C of 25 scholar, and intensity of illumination is 35-40 μm of ol/m-2s-1,
Light application time is that illumination 16 hours, dark are cultivated for 8 hours daily.
Preferably, it in the S6, takes root when operating, chooses the numerous obtained bud of expansion of robust growth, cut in workbench
Base portion callus retains terminal bud part, and long 2-3 cm is seeded on root media, and setting blank control group is taken root
Test, culture of rootage continue 5 weeks, during which observe and record growing state.
Preferably, in the S7, green house control humidity 80% or so, 24-28 DEG C of temperature, 70% natural light of illumination；Adaptability
Training opens bottle cap and carries out hardening, takes out from culture bottle after 3 days and expand numerous obtained bud, and softly clean root under tap water
The culture medium of portion's adhesion, is transplanted in the plastic tub for filling suitable matrix, puts on the polybag there are air hole, pluck after a week
It removes, matrix is made of the rural area soil, peat soil and river sand of 4:3:3, and warm room temperature is 23-27 DEG C, humidity 60-80%, is expanded numerous
Water is watered with when obtained bud transplanting, is poured again when stromal surfaces are slightly dry when subsequent watering etc., to the timely moisturizing of the bud for occurring wilting.
Having the beneficial effect that in the present invention
The present invention is to attempt to cultivate using Plant Biotechnology without Prospect on Kiwifruit Bacterial Canker pathogen " Red Male " in the world for the first time
Kiwi berry seedling can not only lay the foundation for high-quality, disease-free seedlings are mass produced from now on, but also burst to production without Kiwi berry
Other Kiwi berry seedlings of ulcer disease pathogen have good reference function；
The propagation of pathogen is cut off in present invention research from source, and capable of more effectively preventing Prospect on Kiwifruit Bacterial Canker, this is crushing thin
The generation of bacterial diseases, while for having compared and spraying insecticide prevention and treatment, more environmentally-friendly health, compared to for grafting plant, Guo Nongcao
It is more convenient to make management, is conducive to orchard worker's cultivation, under conditions of guaranteeing its interests, due to shoot survival percent height, reproduction speed
Fastly, therefore the yield and quality of Kiwi berry is improved.
Detailed description of the invention
Fig. 1 is a kind of operating process of " Red Male " Kiwi berry without Pathogenic Fungus of Canker seedling fostering technology proposed by the present invention
Figure；
Fig. 2 is for one kind " Red Male " Kiwi berry proposed by the present invention without Pathogenic Fungus of Canker seedling fostering technology without Kiwi berry ulcer
The screening process figure of the bud of sick pathogen.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete
Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.
Referring to Fig.1-2, a kind of " Red Male " Kiwi berry includes the following steps: without Pathogenic Fungus of Canker seedling fostering technology
S1: the Fiber differentiation of " Red Male " Kiwi berry bud takes Chinese gooseberry garden to take branch or blade, by surface sterilization, is training
It supports and induces budding on base；
S2: the proliferation of " Red Male " Kiwi berry bud expands the quantity of bud to the obtained bud of induction into proliferation；
S3: the removing of Prospect on Kiwifruit Bacterial Canker pathogen under anatomical lens, separates shoot tip meristem using the bud of proliferation as material,
It is cultivated to obtain sprouting, and each bud is numbered and is proliferated；
S4: the screening of the bud without Prospect on Kiwifruit Bacterial Canker pathogen, Bacteria Culture identification, by the fertile material of the obtained each bud of S3
It carrying out grinding to be slurried, is coated on bacteria culture media, is cultivated in biochemical cultivation case, observation has abacterial growth, and
It is control with Prospect on Kiwifruit Bacterial Canker pathogen, is compared, filters out the bud of no bacterial growth；PCR identification, with no bacterial growth
Bud extract DNA, and as pcr template, carry out PCR reaction with the special primer of Prospect on Kiwifruit Bacterial Canker pathogen DNA, then right
PCR product is separated by electrophoresis, and whether observation has the specific product of Prospect on Kiwifruit Bacterial Canker pathogen, filters out no specific product
Bud；
S5: expanding the bud of numerous no Prospect on Kiwifruit Bacterial Canker pathogen, to S4 by Bacteria Culture and PCR method screen without macaque
The bud of peach Pathogenic Fungus of Canker is proliferated, and a large amount of bud is obtained；
S6: S5 is expanded numerous obtained bud and is inoculated on root media and carries out by the regeneration without Prospect on Kiwifruit Bacterial Canker pathogen plant
It takes root, to obtain completely without Prospect on Kiwifruit Bacterial Canker pathogen plant；
S7: the regenerated plant of S6 is carried out adaptability by the domestication without Prospect on Kiwifruit Bacterial Canker pathogen plant in being transplanted to greenhouse
Training after one month, moves on to outdoor carry out nature cultivation, forms commercial seedling.
Culture medium includes MS culture medium, 1/2MS culture medium, WPM culture medium and B5 medium, and culture medium is at 121 DEG C
Sterilize 15 min, every liter of 7g containing agar, sucrose 20g；Adjust pH value between 5.8-6.0, wherein 1/2MS culture medium configuration is will to match
The a great number of elements content for setting MS culture medium halves, and every liter of addition sucrose 20g, other contents are identical, being capable of hestening rooting.
In S1, surface sterilization is cleaned once under the tap water that flows slowly with detergent, is rinsed once, is transferred to after sufficiently cleaning
In superclean bench, respectively in the liquor natrii hypochloritis containing 1% effective chlorine and containing 20 and 15 min are sterilized in 0.2% polysorbas20, so
It uses rinsed with sterile water 3 times afterwards.
The condition of culture are as follows: cultivation temperature is 1 DEG C of 25 scholar, and intensity of illumination is 35-40 μm of ol/m-2s-1, light application time is
Illumination 16 hours daily, dark are cultivated for 8 hours.
It in S6, takes root when operating, chooses the numerous obtained bud of expansion of robust growth, base portion callus group is cut in workbench
It knits, retains terminal bud part, long 2-3 cm is seeded on root media, and setting blank control group carries out Rooting, training of taking root
It supports and continues 5 weeks, during which observe and record growing state.
In S7, green house control humidity 80% or so, 24-28 DEG C of temperature, 70% natural light of illumination；Bottle is opened in acclimatization training
Lid carries out hardening, takes out from culture bottle after 3 days and expands numerous obtained bud, and the culture of root adhesion is softly cleaned under tap water
Base is transplanted in the plastic tub for filling suitable matrix, is put on the polybag there are air hole, is extractd after a week, matrix is by 4:3:3
Rural area soil, peat soil and river sand composition, warm room temperature be 23-27 DEG C, humidity 60-80%, expand it is numerous obtain bud transplanting when
It is watered with water, is poured again when stromal surfaces are slightly dry when subsequent watering etc., to the timely moisturizing of the bud for occurring wilting.
The foregoing is only a preferred embodiment of the present invention, but scope of protection of the present invention is not limited thereto,
Anyone skilled in the art in the technical scope disclosed by the present invention, according to the technique and scheme of the present invention and its
Inventive concept is subject to equivalent substitution or change, should be covered by the protection scope of the present invention.
Claims (6)
1. a kind of " Red Male " Kiwi berry is without Pathogenic Fungus of Canker seedling fostering technology, which comprises the steps of:
S1: the Fiber differentiation of " Red Male " Kiwi berry bud takes Chinese gooseberry garden to take branch or blade, by surface sterilization, is training
It supports and induces budding on base；
S2: the proliferation of " Red Male " Kiwi berry bud expands the quantity of bud to the obtained bud of induction into proliferation；
S3: the removing of Prospect on Kiwifruit Bacterial Canker pathogen under anatomical lens, separates shoot tip meristem using the bud of proliferation as material,
It is cultivated to obtain sprouting, and each bud is numbered and is proliferated；
S4: the screening of the bud without Prospect on Kiwifruit Bacterial Canker pathogen, Bacteria Culture identification, by the fertile material of the obtained each bud of S3
It carrying out grinding to be slurried, is coated on bacteria culture media, is cultivated in biochemical cultivation case, observation has abacterial growth, and
It is control with Prospect on Kiwifruit Bacterial Canker pathogen, is compared, filters out the bud of no bacterial growth；PCR identification, with no bacterial growth
Bud extract DNA, and as pcr template, carry out PCR reaction with the special primer of Prospect on Kiwifruit Bacterial Canker pathogen DNA, then right
PCR product is separated by electrophoresis, and whether observation has the specific product of Prospect on Kiwifruit Bacterial Canker pathogen, filters out no specific product
Bud；
S5: expanding the bud of numerous no Prospect on Kiwifruit Bacterial Canker pathogen, to S4 by Bacteria Culture and PCR method screen without macaque
The bud of peach Pathogenic Fungus of Canker is proliferated, and a large amount of bud is obtained；
S6: S5 is expanded numerous obtained bud and is inoculated on root media and carries out by the regeneration without Prospect on Kiwifruit Bacterial Canker pathogen plant
It takes root, to obtain completely without Prospect on Kiwifruit Bacterial Canker pathogen plant；
S7: the regenerated plant of S6 is carried out adaptability by the domestication without Prospect on Kiwifruit Bacterial Canker pathogen plant in being transplanted to greenhouse
Training after one month, moves on to outdoor carry out nature cultivation, forms commercial seedling.
2. one kind " Red Male " Kiwi berry according to claim 1 is without Pathogenic Fungus of Canker seedling fostering technology, feature exists
In the culture medium includes MS culture medium, 1/2MS culture medium, WPM culture medium and B5 medium, and the culture medium is at 121 DEG C
Lower disinfection 15 min, every liter of 7g containing agar, sucrose 20g；Adjust pH value between 5.8-6.0, wherein the configuration of 1/2MS culture medium be by
The a great number of elements content of configuration MS culture medium halves, and every liter of addition sucrose 20g, other contents are identical, being capable of hestening rooting.
3. one kind " Red Male " Kiwi berry according to claim 1 is without Pathogenic Fungus of Canker seedling fostering technology, feature exists
In in the S1, surface sterilization is cleaned once under the tap water that flows slowly with detergent, is rinsed once, is transferred to after sufficiently cleaning super
In net workbench, respectively in the liquor natrii hypochloritis containing 1% effective chlorine and containing 20 and 15 min are sterilized in 0.2% polysorbas20, then
With rinsed with sterile water 3 times.
4. one kind " Red Male " Kiwi berry according to claim 1 is without Pathogenic Fungus of Canker seedling fostering technology, feature exists
In the condition of the culture are as follows: cultivation temperature is 1 DEG C of 25 scholar, and intensity of illumination is 35-40 μm of ol/m-2s-1, light application time is every
Its illumination 16 hours, dark is cultivated for 8 hours.
5. one kind " Red Male " Kiwi berry according to claim 1 is without Pathogenic Fungus of Canker seedling fostering technology, feature exists
In, it in the S6, takes root when operating, chooses the numerous obtained bud of expansion of robust growth, base portion callus is cut in workbench,
Retain terminal bud part, long 2-3 cm is seeded on root media, and setting blank control group carries out Rooting, culture of rootage
Continue 5 weeks, during which observes and records growing state.
6. one kind " Red Male " Kiwi berry according to claim 1 is without Pathogenic Fungus of Canker seedling fostering technology, feature exists
In, in the S7, green house control humidity 80% or so, 24-28 DEG C of temperature, 70% natural light of illumination；Bottle cap is opened in acclimatization training
Hardening is carried out, is taken out from culture bottle after 3 days and expands numerous obtained bud, and softly cleans the culture of root adhesion under tap water
Base is transplanted in the plastic tub for filling suitable matrix, is put on the polybag there are air hole, is extractd after a week, matrix is by 4:3:3
Rural area soil, peat soil and river sand composition, warm room temperature be 23-27 DEG C, humidity 60-80%, expand it is numerous obtain bud transplanting
When be watered with water, poured again when stromal surfaces are slightly dry when subsequent watering etc., to the timely moisturizing of the bud for occurring wilting.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111713410A (en) * | 2020-07-03 | 2020-09-29 | 四川农业大学 | Kiwi explant detoxification method |
| 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 |
-
2019
- 2019-06-12 CN CN201910507729.0A patent/CN110301278A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111713410A (en) * | 2020-07-03 | 2020-09-29 | 四川农业大学 | Kiwi explant detoxification method |
| 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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Application publication date: 20191008 |