CN109618935A - A kind of hybridization paper mulberry efficient in vifro culture method - Google Patents
A kind of hybridization paper mulberry efficient in vifro culture method Download PDFInfo
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- CN109618935A CN109618935A CN201910127376.1A CN201910127376A CN109618935A CN 109618935 A CN109618935 A CN 109618935A CN 201910127376 A CN201910127376 A CN 201910127376A CN 109618935 A CN109618935 A CN 109618935A
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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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- Developmental Biology & Embryology (AREA)
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Abstract
A kind of hybridization paper mulberry efficient in vifro culture method, the propagation method including the following steps: (1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA, it is spare after sterilizing 21 minutes；(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；(4) explant rinses；(5) explant sterilizes；(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and blade, removing stipule of bleaching, one sprout of every bottle of inoculation；(7) it cultivates.The constituent and illumination condition of culture medium carry out reasonably optimizing, improve tissue culture efficiency, inductivity, breeding potential and rooting rate with higher.
Description
Technical field
The present invention relates to paper mulberry planting technology field more particularly to a kind of hybridization paper mulberry efficient in vifro culture method.
Background technique
Hybridization paper mulberry is Moraceae paper mulberry platymiscium, is that the comprehensive means such as China's last decade space treatment space breeding are cultivated
Space flight out hybridizes paper mulberry, has the characteristics such as fast-growing, high yield, high-quality, how anti-.Paper mulberry leaf contains crude protein and flavonoids, is one
The good plant protein fodder of kind, comprehensive nutrient ingredient are more than bean, and dregs of beans, fish meal can be replaced to support for various animals
It grows.
Hybridization paper mulberry seedling raising manners have grow directly from seeds branch cutting nursery and tissue-culturing rapid propagation nursery.But cuttage and seedling culture breeds coefficient
Low, the breeding cycle is long, and survival rate is low, it is difficult to meet the needs of current commerial growing and popularization.Tissue-culturing rapid propagation nursery then can be
A large amount of breed good strains in a short time, and be not subject to seasonal restrictions.However, cultivating material in existing hybridization paper mulberry group culturation rapid propagating technology
Material there are rooting rates it is low, survival rate is low the problems such as.
Summary of the invention
For above-mentioned deficiency in the prior art, the present invention provides a kind of hybridization paper mulberry efficient in vifro culture method, trainings
The constituent and illumination condition for supporting base carry out reasonably optimizing, improve tissue culture efficiency, inductivity with higher, breeding potential
And rooting rate.
In order to achieve the above object of the invention, the present invention use the specific scheme is that
A kind of hybridization paper mulberry efficient in vifro culture method, the propagation method including the following steps:
(1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA goes out
It is spare after bacterium 21 minutes；
(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；
(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；
(4) explant rinses: terminal bud being subtracted into the stem section or the wooden stem section of resting bud band part of 2-2.5cm, is first rushed with tap water
It washes 15-20 minutes, the pollutant on explant surface is gently brushed off with soft hairbrush；
(5) explant sterilizes: cleaned explant being put into after sterilizing 15s in 75% alcohol and is transferred to ready sterile water
In, it is transferred to after jiggling 1 minute in the mercuric chloride solution that 1-2 drop dish washing liquid 0.1g/L is added, disinfection was transferred to nothing after 15 minutes
It is cleaned 3-4 times in bacterium water, obtains the explant disinfected；
(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and leaf of bleaching
Piece removes stipule, one sprout of every bottle of inoculation；
(7) it cultivates: under illumination 3000-4000LX, 27-30 DEG C of temperature, the gnotobasis of humidity 60%-80%, in the medium
Culture 35-45 days selects the sprout normally sprouted and transfers as female bottle except the part that do not sprout seriously with browning of depolluting
Expand numerous.
The invention has the benefit that
The present invention selects hybridization paper mulberry terminal bud or full resting bud as explant, and meristematic capacity is strong, is easier to division and reproduction；Light
According to 3000-4000LX, 27-30 DEG C of temperature, the gnotobasis of humidity 60%-80%, its environmental suitability is improved, and culture medium
Constituent and illumination condition carry out reasonably optimizing, improve tissue culture efficiency, inductivity with higher, breeding potential and take root
Rate carries out the nursery of hybridization paper mulberry in this method using tissue culture rapid propagation in vitro, and breeding efficiency is high, and breeding coefficient is big, reduces and educates
Seedling cost, and the in vitro seedling of gained tissue culture is healthy and strong, quality is high.
Specific embodiment
The present invention is further described below by way of specific embodiment, but the present invention is not limited only to following embodiment.At this
In the range of invention or the contents of the present invention are not being departed from, in spirit and scope, the change that carries out to the present invention is combined or replaced
It changes, will be apparent to the person skilled in the art, and be included within the scope of the present invention.
Embodiment 1: a kind of hybridization paper mulberry efficient in vifro culture method, the propagation method including the following steps:
(1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA goes out
It is spare after bacterium 21 minutes；
(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；
(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；
(4) explant rinses: terminal bud being subtracted into the stem section or the wooden stem section of resting bud band part of 2cm, first rinses 15 with tap water
Minute, the pollutant on explant surface is gently brushed off with soft hairbrush；
(5) explant sterilizes: cleaned explant being put into after sterilizing 15s in 75% alcohol and is transferred to ready sterile water
In, it is transferred to after jiggling 1 minute in the mercuric chloride solution that 1-2 drop dish washing liquid 0.1g/L is added, disinfection was transferred to nothing after 15 minutes
It is cleaned 3-4 times in bacterium water, obtains the explant disinfected；
(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and leaf of bleaching
Piece removes stipule, one sprout of every bottle of inoculation；
(7) it cultivates: under illumination 3000LX, 27 DEG C of temperature, the gnotobasis of humidity 60%, cultivating 35 days, remove in the medium
Pollution and the part do not sprouted seriously of browning, select the sprout that normally sprouts as female bottle carry out switching expand it is numerous.
Embodiment 2: a kind of hybridization paper mulberry efficient in vifro culture method, the propagation method including the following steps:
(1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA goes out
It is spare after bacterium 21 minutes；
(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；
(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；
(4) explant rinses: terminal bud being subtracted into the stem section or the wooden stem section of resting bud band part of 2.3cm, is first rinsed with tap water
18 minutes, the pollutant on explant surface is gently brushed off with soft hairbrush；
(5) explant sterilizes: cleaned explant being put into after sterilizing 15s in 75% alcohol and is transferred to ready sterile water
In, it is transferred to after jiggling 1 minute in the mercuric chloride solution that 1-2 drop dish washing liquid 0.1g/L is added, disinfection was transferred to nothing after 15 minutes
It is cleaned 3-4 times in bacterium water, obtains the explant disinfected；
(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and leaf of bleaching
Piece removes stipule, one sprout of every bottle of inoculation；
(7) it cultivates: under illumination 3500LX, 29 DEG C of temperature, the gnotobasis of humidity 70%, cultivating 40 days, remove in the medium
Pollution and the part do not sprouted seriously of browning, select the sprout that normally sprouts as female bottle carry out switching expand it is numerous.
Embodiment 3: a kind of hybridization paper mulberry efficient in vifro culture method, the propagation method including the following steps:
(1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA goes out
It is spare after bacterium 21 minutes；
(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；
(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；
(4) explant rinses: terminal bud being subtracted into the stem section or the wooden stem section of resting bud band part of 2.5cm, is first rinsed with tap water
20 minutes, the pollutant on explant surface is gently brushed off with soft hairbrush；
(5) explant sterilizes: cleaned explant being put into after sterilizing 15s in 75% alcohol and is transferred to ready sterile water
In, it is transferred to after jiggling 1 minute in the mercuric chloride solution that 1-2 drop dish washing liquid 0.1g/L is added, disinfection was transferred to nothing after 15 minutes
It is cleaned 3-4 times in bacterium water, obtains the explant disinfected；
(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and leaf of bleaching
Piece removes stipule, one sprout of every bottle of inoculation；
(7) it cultivates: under illumination 4000LX, 30 DEG C of temperature, the gnotobasis of humidity 80%, cultivating 45 days, remove in the medium
Pollution and the part do not sprouted seriously of browning, select the sprout that normally sprouts as female bottle carry out switching expand it is numerous.
The above description is only a preferred embodiment of the patent of the present invention, is not intended to limit the invention patent, all at this
Made any modifications, equivalent replacements, and improvements etc., should be included in the invention patent within the spirit and principle of patent of invention
Protection scope within.
Claims (1)
1. a kind of hybridization paper mulberry efficient in vifro culture method, which is characterized in that the propagation method including the following steps:
(1) preparation of culture medium: medium component: MS+5.8g/L agar+30g/L sugar+1.0mg/L6-BA+0.05mg/LNAA goes out
It is spare after bacterium 21 minutes；
(2) prepared by aqua sterilisa: taking 2/3 pure water to sterilize with 250ml culture bottle, to obtain sterile water within 21 minutes spare；
(3) prepared by explant: taking healthy and strong hybridization paper mulberry terminal bud or full resting bud as inoculation material；
(4) explant rinses: terminal bud being subtracted into the stem section or the wooden stem section of resting bud band part of 2-2.5cm, is first rushed with tap water
It washes 15-20 minutes, the pollutant on explant surface is gently brushed off with soft hairbrush；
(5) explant sterilizes: cleaned explant being put into after sterilizing 15s in 75% alcohol and is transferred to ready sterile water
In, it is transferred to after jiggling 1 minute in the mercuric chloride solution that 1-2 drop dish washing liquid 0.1g/L is added, disinfection was transferred to nothing after 15 minutes
It is cleaned 3-4 times in bacterium water, obtains the explant disinfected；
(6) explant is inoculated with: the explant disinfected being cut into the segment of 1cm or so, cuts off the wound part and leaf of bleaching
Piece removes stipule, one sprout of every bottle of inoculation；
(7) it cultivates: under illumination 3000-4000LX, 27-30 DEG C of temperature, the gnotobasis of humidity 60%-80%, in the medium
Culture 35-45 days selects the sprout normally sprouted and transfers as female bottle except the part that do not sprout seriously with browning of depolluting
Expand numerous.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201910127376.1A CN109618935A (en) | 2019-02-20 | 2019-02-20 | A kind of hybridization paper mulberry efficient in vifro culture method |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113100064A (en) * | 2021-05-10 | 2021-07-13 | 刘尚文 | Culture medium for tissue culture of broussonetia papyrifera and preparation method |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108338071A (en) * | 2017-12-29 | 2018-07-31 | 天长市金农农业发展有限公司 | A kind of asexual multiplication seedling method of hybridization paper mulberry |
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2019
- 2019-02-20 CN CN201910127376.1A patent/CN109618935A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108338071A (en) * | 2017-12-29 | 2018-07-31 | 天长市金农农业发展有限公司 | A kind of asexual multiplication seedling method of hybridization paper mulberry |
Non-Patent Citations (1)
| Title |
|---|
| 万文等: "杂交构树茎段组培快繁体系的建立", 《福建林业科技》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113100064A (en) * | 2021-05-10 | 2021-07-13 | 刘尚文 | Culture medium for tissue culture of broussonetia papyrifera and preparation method |
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