CN109380084A - A kind of tissue cultivation rapid breeding method of the seed of feather cockscomb - Google Patents
A kind of tissue cultivation rapid breeding method of the seed of feather cockscomb Download PDFInfo
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- CN109380084A CN109380084A CN201811410521.9A CN201811410521A CN109380084A CN 109380084 A CN109380084 A CN 109380084A CN 201811410521 A CN201811410521 A CN 201811410521A CN 109380084 A CN109380084 A CN 109380084A
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- feather cockscomb
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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
-
- A—HUMAN NECESSITIES
- 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
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
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- A—HUMAN NECESSITIES
- 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
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
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- A—HUMAN NECESSITIES
- 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
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
-
- 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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- Environmental Sciences (AREA)
- Developmental Biology & Embryology (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Soil Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Inorganic Chemistry (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The problems such as the invention discloses a kind of tissue cultivation rapid breeding method of seed of feather cockscomb, seed of feather cockscomb seedling is mainly bred by seed and cuttage mode, and there are the period is long, at high cost, inefficiency.Therefore, the present invention is using seed of feather cockscomb stem-segment with node as explant, by adventitious bud inducing, be proliferated, take root, seed of feather cockscomb plant again in vitro has successfully been obtained in the processes such as acclimatization and transplants, establish seed of feather cockscomb tissue culture rapid propagation technique system, for carrying out quickly breeding and be widely applied for seed of feather cockscomb excellent variety, the industrialized development of promotion is of great significance.
Description
Technical field
The present invention relates to the methods of Plant Tissue Breeding in agricultural biotechnologies, specifically, being related to a kind of seed of feather cockscomb
Tissue cultivation rapid breeding method.
Background technique
The seed of feather cockscomb is amaranthaceous plant, takes the dry mature seed of feather cockscomb.Main product Guangdong, Guangxi, Fujian, Shandong.General 7 ~ 9
Harvesting complete stool is dried when moon seed maturation, lays seed.It is lightly seasoned without gas.With clearing liver and improving vision, screen is moved back.It is raw for red eye, swell pain
The effects of screen film, eye-blurred.
Currently, seed of feather cockscomb seedling is mainly bred by seed and cuttage mode, although seed sowing can obtain in a short time
A large amount of seedling are obtained, but because the seed of feather cockscomb is monoecism cross-pollinatd plant, offspring's Yi Fasheng trait segregation, inhereditary feature shakiness
It is fixed, the merit of parent easy to be lost.And cuttage mode breeds and needs a large amount of branch, and needs 2 from cuttage to transplanting crop field
Month, it takes a long time.Therefore, alleviate its seedling tension pressure using plant tissue culture technique, carry out Sustainable Development and Utilization
It is very necessary.Therefore, the present invention is using seed of feather cockscomb stem-segment with node as explant, by adventitious bud inducing, is proliferated, takes root, refines
Seed of feather cockscomb plant again in vitro has successfully been obtained in the processes such as transplantation of seedlings, establishes seed of feather cockscomb tissue culture rapid propagation technique system, right
There is important meaning in the quick industrialized development bred and be widely applied, promote the seed of feather cockscomb for carrying out seed of feather cockscomb excellent variety
Justice.
Summary of the invention
The purpose of the present invention is to provide a kind of tissue cultivation rapid breeding methods of seed of feather cockscomb, by evoking adventive bud, grow thickly
Seed of feather cockscomb plant again in vitro has successfully been obtained in the stages such as bud proliferation, rooting of vitro seedling, acclimatization and transplants, establishes seed of feather cockscomb tissue cultures
Quick breeding technology system, to realize the purpose of the present invention.
A kind of tissue cultivation rapid breeding method of seed of feather cockscomb of the invention, including processing step below:
Step (1), evoking adventive bud: selection seed of feather cockscomb current year green tape section rattan was explant, through 75% alcohol disinfecting 5~20 seconds
After kind, it is placed in 0.1% mercuric chloride solution and is sterilized 34 minutes with aseptic water washing 9 times, then used aseptic water washing 9 times, through nothing
The belt segment rattan of 6cm or so is cut into after bacterium filter paper suck dry moisture, and is inoculated into induced medium, under the conditions of 25~28 DEG C
Full dark culture, until induced synthesis adventitious bud；Induced medium are as follows: MS+0.5mg/LTDZ+2.3mg/L 6-BA+0.8mg/
LNAA+30g/L sucrose+6.5g/L agar, pH 5.7；
Step (2), squamous subculture: by step (1) obtain adventitious bud cut from base portion, be inoculated on proliferated culture medium carry out after
It is commissioned to train feeding, inoculation is placed on daily illumination 17 hours, intensity of illumination 2400lx, and cultivation temperature is trained under conditions of being 25~28 DEG C
It supports, switching in 34 days is primary；Proliferated culture medium are as follows: MS+2.3mg/L 6-BA+0.9mg/LNAA+33g/L sucrose+6.6g/L agar,
PH is 5.9；
Culture of rootage: the long adventitious bud to 6cm of step (2) squamous subculture is cut and is inoculated on root media for step (3)
Root induction is carried out, it is first dark culture 7 days full under the conditions of 25~28 DEG C after inoculation, it is subsequently placed in daily illumination 17 hours, illumination
Intensity is 3400lx, and cultivation temperature is cultivated under conditions of being 25~28 DEG C to taking root；Root media are as follows: 1/2MS+0.9mg/
LIBA+2.3mg/L GGR+33g/L sucrose+6.6g/L agar, pH 5.9；
Acclimatization and transplants: the culture bottle cap opening of the long rooting tube plantlet to 12cm is placed under natural lighting and is refined for step (4)
After seedling 9 days, test tube seedling is taken out from culture bottle, washes off root culture medium, is planted by peat soil: farmyard manure: river sand (8:2:2)
In the matrix being mixed into and it is colonized in big Tanaka.
The invention has the advantages that the present invention using seed of feather cockscomb stem-segment with node as explant, passes through adventitious bud inducing, proliferation, life
Seed of feather cockscomb plant again in vitro has successfully been obtained in the processes such as root, acclimatization and transplants, establishes seed of feather cockscomb tissue culture rapid propagation technique body
System, the industrialized development quickly bred and be widely applied, promote the seed of feather cockscomb for carrying out seed of feather cockscomb excellent variety have heavy
Want meaning.Tissue cultivation rapid breeding method of the present invention has the characteristics that simple, easy, economical.What is be bred as through the invention utilizes plant
Object tissue culture technique carries out the large-scale production of seed of feather cockscomb seedling, establishes tissue-cultured seedling transplanting survival rate and reaches 98% or more, can be with
High quality seedling guarantee is provided for seed of feather cockscomb large-scale planting.
Specific embodiment
It the following examples are further illustrations of the invention, is not limitation of the present invention.
Embodiment 1:
(1) evoking adventive bud: selection seed of feather cockscomb current year green tape section rattan is explant, after 75% alcohol disinfecting, 15 seconds kinds, is used
It is placed in 0.1% mercuric chloride solution for aseptic water washing 5 times and to sterilize 18 minutes, then use aseptic water washing 9 times, through aseptic filter paper
The belt segment rattan of 7cm or so is cut into after suck dry moisture, and is inoculated into induced medium, the full dark culture 21 under the conditions of 25 DEG C
It can induced synthesis adventitious bud, pollution rate is down to 6% hereinafter, adventitious bud induction frequency is up to 93.6%.The induced medium
Are as follows: MS+0.2mg/LTDZ+1.2mg/L 6-BA+0.5mg/LNAA+20g/L sucrose+6.8g/L agar, pH 5.6;
(2) squamous subculture: the adventitious bud that step (1) obtains is cut from base portion, is inoculated into progress subculture training on proliferated culture medium
It supports, inoculation is placed on daily illumination 15 hours, intensity of illumination 1900lx, and cultivation temperature is cultivated under conditions of being 25 DEG C, and 24 days
Switching is primary, growth coefficient 7.9.The proliferated culture medium are as follows: MS+1.5mg/L 6-BA+0.2mg/LNAA+25g/L sugarcane
Sugar+6.6g/L agar, pH 5.6；
(3) culture of rootage: the adventitious bud that step (2) squamous subculture is grown to 7cm, which is cut and is inoculated on root media, to carry out
Root induction, it is first dark culture 5 days full under the conditions of 25 DEG C after inoculation, it is subsequently placed in daily illumination 16 hours, intensity of illumination is
2900lx, cultivation temperature is cultivated 12 days under conditions of being 25 DEG C to be started to take root, and rooting rate is up to 97%.The culture of rootage
Base are as follows: 1/2MS+0.3mg/LIBA+1.0mg/L GGR+19g/L sucrose+6.6g/L agar, pH 5.6.
(4) acclimatization and transplants: the culture bottle cap opening of the long rooting tube plantlet to 12cm is placed under natural lighting and is refined
After seedling 8 days, test tube seedling is taken out from culture bottle, washes off root culture medium, is planted by peat soil: farmyard manure: river sand (8:2:2)
In the matrix being mixed into and it is colonized in big Tanaka, transplanting survival rate 100%.
Embodiment 2:
(1) evoking adventive bud: selection seed of feather cockscomb current year green tape section rattan is explant, after 75% alcohol disinfecting, 19 seconds kinds, with nothing
Bacterium water is rinsed to be placed in 0.1% mercuric chloride solution for 6 times and sterilize 19 minutes, then with aseptic water washing 8 times, is inhaled through aseptic filter paper
The belt segment rattan of 6cm or so is cut into after solid carbon dioxide point, and is inoculated into induced medium, full dark culture 16 days under the conditions of 26 DEG C
Can induced synthesis adventitious bud, pollution rate is down to 8% hereinafter, adventitious bud induction frequency is up to 96.5%.The induced medium are as follows:
MS+0.4mg/LTDZ+1.9mg/L 6-BA+1.5mg/LNAA+30g/L sucrose+5.5g/L agar, pH 5.8.
(2) squamous subculture: by step (1) obtain adventitious bud cut from base portion, be inoculated on proliferated culture medium carry out after
It is commissioned to train feeding, inoculation is placed on daily illumination 17 hours, intensity of illumination 2400lx, and cultivation temperature is cultivated under conditions of being 26 DEG C,
Switching in 28 days is primary, growth coefficient 9.5.The proliferated culture medium are as follows: MS+1.0mg/L 6-BA+0.5mg/LNAA+35g/
L sucrose+6.5g/L agar, pH 5.6.
(3) culture of rootage: the long adventitious bud to 6cm of step (2) squamous subculture is cut and is inoculated on root media
Root induction is carried out, it is first dark culture 6 days full under the conditions of 26 DEG C after inoculation, it is subsequently placed in daily illumination 18 hours, intensity of illumination
For 3400lx, cultivation temperature is cultivated 16 days under conditions of being 26 DEG C to be started to take root, and rooting rate is up to 98%.The training of taking root
Support base are as follows: 1/2MS+1.0mg/LIBA+2.0mg/L GGR+21g/L sucrose+5.0g/L agar, pH 5.6.
(4) acclimatization and transplants: the culture bottle cap opening of the long rooting tube plantlet to 12cm is placed under natural lighting and is refined
After seedling 6 days, test tube seedling is taken out from culture bottle, washes off root culture medium, is planted by peat soil: farmyard manure: river sand (8:2:2)
In the matrix being mixed into and it is colonized in big Tanaka, transplanting survival rate 100%.
Claims (1)
1. a kind of tissue cultivation rapid breeding method of the seed of feather cockscomb, it is characterised in that comprise the following steps that:
Step (1), evoking adventive bud: selection seed of feather cockscomb current year green tape section rattan was explant, through 75% alcohol disinfecting 5~20 seconds
After kind, it is placed in 0.1% mercuric chloride solution and is sterilized 34 minutes with aseptic water washing 9 times, then used aseptic water washing 9 times, through nothing
The belt segment rattan of 6cm or so is cut into after bacterium filter paper suck dry moisture, and is inoculated into induced medium, under the conditions of 25~28 DEG C
Full dark culture, until induced synthesis adventitious bud；Induced medium are as follows: MS+0.5mg/LTDZ+2.3mg/L 6-BA+0.8mg/
LNAA+30g/L sucrose+6.5g/L agar, pH 5.7；
Step (2), squamous subculture: by step (1) obtain adventitious bud cut from base portion, be inoculated on proliferated culture medium carry out after
It is commissioned to train feeding, inoculation is placed on daily illumination 17 hours, intensity of illumination 2400lx, and cultivation temperature is trained under conditions of being 25~28 DEG C
It supports, switching in 34 days is primary；Proliferated culture medium are as follows: MS+2.3mg/L 6-BA+0.9mg/LNAA+33g/L sucrose+6.6g/L agar,
PH is 5.9；
Culture of rootage: the long adventitious bud to 6cm of step (2) squamous subculture is cut and is inoculated on root media for step (3)
Root induction is carried out, it is first dark culture 7 days full under the conditions of 25~28 DEG C after inoculation, it is subsequently placed in daily illumination 17 hours, illumination
Intensity is 3400lx, and cultivation temperature is cultivated under conditions of being 25~28 DEG C to taking root；Root media are as follows: 1/2MS+0.9mg/
LIBA+2.3mg/L GGR+33g/L sucrose+6.6g/L agar, pH 5.9；
Acclimatization and transplants: the culture bottle cap opening of the long rooting tube plantlet to 12cm is placed under natural lighting and is refined for step (4)
After seedling 9 days, test tube seedling is taken out from culture bottle, washes off root culture medium, is planted by peat soil: farmyard manure: river sand (8:2:2)
In the matrix being mixed into and it is colonized in big Tanaka.
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| CN201811410521.9A CN109380084A (en) | 2018-11-23 | 2018-11-23 | A kind of tissue cultivation rapid breeding method of the seed of feather cockscomb |
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| CN201811410521.9A CN109380084A (en) | 2018-11-23 | 2018-11-23 | A kind of tissue cultivation rapid breeding method of the seed of feather cockscomb |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082138A (en) * | 2014-06-28 | 2014-10-08 | 玉林师范学院 | Tissue-culture rapid propagation method of Aristolochia fordiana Hemsl |
| CN104686345A (en) * | 2015-02-24 | 2015-06-10 | 陈桂容 | Tissue culture rapid propagation method of liquorice |
| CN104798684A (en) * | 2015-04-25 | 2015-07-29 | 玉林师范学院 | Tissue culture rapid propagation method for plukenetia volubilis L. |
-
2018
- 2018-11-23 CN CN201811410521.9A patent/CN109380084A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104082138A (en) * | 2014-06-28 | 2014-10-08 | 玉林师范学院 | Tissue-culture rapid propagation method of Aristolochia fordiana Hemsl |
| CN104686345A (en) * | 2015-02-24 | 2015-06-10 | 陈桂容 | Tissue culture rapid propagation method of liquorice |
| CN104798684A (en) * | 2015-04-25 | 2015-07-29 | 玉林师范学院 | Tissue culture rapid propagation method for plukenetia volubilis L. |
| CN104798684B (en) * | 2015-04-25 | 2017-04-05 | 玉林师范学院 | A kind of tissue cultivation rapid breeding method of astral oil rattan |
Non-Patent Citations (1)
| Title |
|---|
| 杨冬业 等: "青葙高效再生体系的建立", 《作物杂志》 * |
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Application publication date: 20190226 |