CN105993937A - Producing method for Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant - Google Patents
Producing method for Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant Download PDFInfo
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- CN105993937A CN105993937A CN201610312866.5A CN201610312866A CN105993937A CN 105993937 A CN105993937 A CN 105993937A CN 201610312866 A CN201610312866 A CN 201610312866A CN 105993937 A CN105993937 A CN 105993937A
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- poplar
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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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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention belongs to the technical field of plant tissue culture, and particularly relates to a producing method for a Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant. The producing method for the Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant includes the steps of selecting and disinfecting explants, inducing callus tissue, inducing adventitious buds and inducing roots. The producing method is short in culture cycle, little in variation and easy to operate, culture conditions can be controlled manually, the foundation is laid for genetic transformation and breed improvement with Lux poplar and Cathay poplar hybrid F1 superior individual plants serving as good scientific research materials, and therefore a good variety resistant to saline and alkaline soil is provided for large-area production of a poplar artificial forest.
Description
One technical field
The invention belongs to field of plant tissue culture technique, be specifically related to a kind of Lu Kesi poplar and Cathay poplar F-1 hybrids fine individual plant leaf
The production method of sheet regeneration plant.
Two technical backgrounds
Willow because of its fast growth, world's distribution is wide, artificial forest planting area big, have become as 21 century and improve human survival
Environment, meets one of most potential seeds of human lives's demand.Have become as the moulds such as world's xylophyta breeding, cultivation at present
Formula seeds.
In recent years, Poplar Plantation area continuous enlargement, the problem occurring in that usable land inadequate resource, and there is big face in China
Long-pending saline and alkaline beach ground, area reaches 0.27 hundred million hm2, how to strengthen the utilization to saline and alkaline beach ground, become in Poplar Breeding and close
The focus of note.
Three summary of the invention
The present invention needs the problem solved to be for the demand in willow fine germplasm resources Salt And Alkali Tolerance breeding, has invented a kind of Shandong gram
The production method of this poplar and Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant, for willow fine germplasm resources genetic transformation and
Breed improvement quickly provides research material.
The production method of Lu Kesi poplar of the present invention and Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant is by following steps structure
Become:
(1) selection of outer implant and sterilization:
The young leaflet tablet of the healthy and strong disease-free Lu Kesi poplar of growth selection and Cathay poplar F-1 hybrids fine individual plant is as outer implant, by material
Expect to rinse 1h through flowing water, remove surface irregularities, on aseptic manipulation work table, first with the aseptic water washing 3 times of sterilizing, then use
75% dehydrated alcohol sterilization 30s, then uses aseptic water washing 3 times, then disinfects 5-7min with 0.1% mercuric chloride,
Afterwards with the aseptic water washing 4 times of sterilizing；
(2) induction of callus:
On superclean bench, the blade disinfected is cut into the square of 6mm size, is inoculated in callus induction culture medium and (joins
Fang Wei: MS+0.5mg/L 6-BA+0.15mg/L NAA+30g/L sucrose+7g/L agar；Ph is adjusted to 5.8), the back side of leaf is tight
Patch culture medium, every bottle graft kind 3-4 block, induce illumination condition 12h/d, 2000lux, temperature 23-28 DEG C；
(3) induction of adventitious bud:
Yellow green callus after successive transfer culture 1 time is transferred on evoking adventive bud division culture medium (formula is:
MS+0.5mg/L 6-BA+0.05mg/L NAA+30g/L sucrose+7g/L agar；Ph is adjusted to 5.8), induce illumination condition 12h/d,
Intensity of illumination about 2000lux, temperature 23-28 DEG C；
(4) induction of root:
Obtaining regeneration bud through Induce aerosor, bud to be regenerated grows to 4-5 sheet leaf, and length 0.8-1.2cm Gao Shike proceeds to raw
(formula is: 1/2MS+0.6mg/LIBA+0.2mg/L NAA+30g/L sucrose+7g/L agar in root culture medium root induction；Ph adjusts
It is 5.8), induce illumination condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C；.
Lu Kesi poplar of the present invention and Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant cultivation cycle are 45-55 days.
Compared with prior art, it provides the benefit that the present invention: the present invention uses Lu Kesi poplar and Cathay poplar F-1 hybrids fine individual plant
Blade, as material, is drawn materials conveniently, abundant amount, and after inoculation, within a short period of time forms callus until taking root and forming seedling
Plant, gained plant strain growth is healthy and strong, and average root length all significantly increases.Breeding coefficient is about 8.6, and condition of culture can artificially be controlled
System.Utilize this method, Lu Kesi poplar and the seedling breeding coefficient of Cathay poplar F-1 hybrids fine individual plant and survival rate, training can be improved
The cycle of supporting is short, and variation is few, condition of culture can manual control, operational approach is simple, excellent with Cathay poplar F-1 hybrids for Lu Kesi poplar
Individual plant carries out the research such as genetic transformation and breed improvement as excellent scientific research material and lays the foundation, thus is big of Poplar Plantation
The long-pending improved seeds that offer Salt And Alkali Tolerance ground is provided.
Four accompanying drawing explanations
Fig. 1 Lu Kesi poplar and the leaf explant of Cathay poplar F-1 hybrids fine individual plant regeneration plant
Fig. 2 Lu Kesi poplar and the callus of Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant
Fig. 3 Lu Kesi poplar and the Multiple Buds of Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant
Fig. 4 Lu Kesi poplar and the rooted plantlet of Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant
Five detailed description of the invention
1, the selection of propagating materials
Test material takes from the Lu Kesi poplar on Jinling School of Science and Technology willow base and hybridization Cathay poplar F-1 hybrids fine individual plant.
2, the sterilization of propagating materials
Material is rinsed 1h through flowing water and removes surface irregularities, on aseptic manipulation work table, first use aseptic water washing 3 times, then use
75% dehydrated alcohol sterilization 30s, then aseptic water washing 3 times, then disinfect 5-7min, finally with 0.1% mercuric chloride
With the aseptic water washing 4 times of sterilizing；See accompanying drawing 1.
3, the induction of callus
On superclean bench, the blade disinfected is cut into the square of 6mm size, is inoculated on calli induction media, leaf
The back side be close to culture medium, every bottle graft kind 3-4 block, induce illumination condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C；
See accompanying drawing 2
4, the induction of adventitious bud
Yellow green callus after continuous successive transfer culture 2-3 time is transferred on evoking adventive bud division culture medium, induction
Illumination condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C；See accompanying drawing 3
5, the induction of root
Obtaining regeneration bud through Induce aerosor, bud to be regenerated grows to 4-5 sheet leaf, and length 1cm Gao Shike proceeds to root culture
Base root induction, induces illumination condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C.Seedling takes root situation, sees
Accompanying drawing 4.
Claims (2)
1. Yi Zhong Lu Kesi poplar and the production method of Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant, is characterized in that by following steps
Constitute:
(1) selection of outer implant and sterilization
The young leaflet tablet of the healthy and strong disease-free Lu Kesi poplar of growth selection and Cathay poplar F-1 hybrids fine individual plant is as outer implant, by material
Expect to rinse 1h through flowing water, remove surface irregularities, on aseptic manipulation work table, first with the aseptic water washing 3 times of sterilizing, then use
75% dehydrated alcohol sterilization 30s, then uses aseptic water washing 3 times, then disinfects 5-7min, finally with 0.1% mercuric chloride
Rinse 4 times with sterilized water concussion；
2) induction of callus
On superclean bench, the blade disinfected is cut into the square of 6mm size, is inoculated in callus induction culture medium, training
The formula supporting base is: the NAA+30g/L sucrose+7g/L agar of the 6-BA+0.15mg/L of MS+0.5mg/L；Ph is adjusted to 5.8,
Culture medium, every bottle graft kind 3-4 block are close in the back side of leaf, induce illumination condition 12h/d, intensity of illumination 2000lux, temperature
23-28℃；
3) induction of adventitious bud
Yellow green callus after successive transfer culture 1 time is transferred on evoking adventive bud division culture medium, joining of culture medium
NAA+30g/L sucrose+7g/L the agar of the 6-BA+0.05mg/L of Fang Wei: MS+0.5mg/L；Ph is adjusted to 5.8, induces illumination
Condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C；
4) induction of root
Obtaining regeneration bud through Induce aerosor, bud to be regenerated grows to 4-5 sheet leaf, proceeds to take root during length 0.8-1.2cm height
Culture medium root induction, the formula of culture medium is: the NAA+30g/L sucrose+7g/L of the IBA+0.2mg/L of 1/2MS+0.6mg/L
Agar；Ph is adjusted to 5.8, induces illumination condition 12h/d, intensity of illumination 2000lux, temperature 23-28 DEG C.
A kind of Lu Kesi poplar and the production method of Cathay poplar F-1 hybrids fine individual plant leaf regeneration plant,
It is characterized in that breeding coefficient is 8.6, cultivation cycle is 45-55 days.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610312866.5A CN105993937A (en) | 2016-05-12 | 2016-05-12 | Producing method for Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant |
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| CN201610312866.5A CN105993937A (en) | 2016-05-12 | 2016-05-12 | Producing method for Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant |
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| CN201610312866.5A Pending CN105993937A (en) | 2016-05-12 | 2016-05-12 | Producing method for Lux poplar and Cathay poplar hybrid F1 superior individual plant leaf regeneration plant |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508677A (en) * | 2016-11-03 | 2017-03-22 | 金陵科技学院 | Method for producing populus euphratica and populus talassica hybrid F1-generation excellent plant leave regeneration plant |
| CN106561458A (en) * | 2016-11-09 | 2017-04-19 | 金陵科技学院 | Production method for regenerated plants of good single plant leaves of hybrid F1 generation of populus deltoids 69 and populus euramericana 72 |
| CN107114235A (en) * | 2017-04-10 | 2017-09-01 | 中国林业科学研究院林业研究所 | A kind of method that utilization DNA methylation inhibitor builds plant population |
| CN111543318A (en) * | 2020-01-19 | 2020-08-18 | 内蒙古农业大学 | Sterile micro-cuttage rapid propagation method for populus deltoides |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101695279A (en) * | 2009-09-11 | 2010-04-21 | 湖南茂源林业有限责任公司 | Method for tissue culture of populus deltoids forest 101 |
| CN104396754A (en) * | 2014-11-25 | 2015-03-11 | 东北林业大学 | Populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method |
| KR20150093517A (en) * | 2014-02-07 | 2015-08-18 | 충북대학교 산학협력단 | Method for mass propagation of callus and method for mass division of Populus euramericana utilizing stem segments of Populus euramericana |
-
2016
- 2016-05-12 CN CN201610312866.5A patent/CN105993937A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101695279A (en) * | 2009-09-11 | 2010-04-21 | 湖南茂源林业有限责任公司 | Method for tissue culture of populus deltoids forest 101 |
| KR20150093517A (en) * | 2014-02-07 | 2015-08-18 | 충북대학교 산학협력단 | Method for mass propagation of callus and method for mass division of Populus euramericana utilizing stem segments of Populus euramericana |
| CN104396754A (en) * | 2014-11-25 | 2015-03-11 | 东北林业大学 | Populus trichocarpa Torr.Gray leaf adventitious bud induction and plant regeneration method |
Non-Patent Citations (1)
| Title |
|---|
| 程明珠: "杨树优良无性系再生体系的建立及转Bt基因的研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508677A (en) * | 2016-11-03 | 2017-03-22 | 金陵科技学院 | Method for producing populus euphratica and populus talassica hybrid F1-generation excellent plant leave regeneration plant |
| CN106561458A (en) * | 2016-11-09 | 2017-04-19 | 金陵科技学院 | Production method for regenerated plants of good single plant leaves of hybrid F1 generation of populus deltoids 69 and populus euramericana 72 |
| CN107114235A (en) * | 2017-04-10 | 2017-09-01 | 中国林业科学研究院林业研究所 | A kind of method that utilization DNA methylation inhibitor builds plant population |
| CN111543318A (en) * | 2020-01-19 | 2020-08-18 | 内蒙古农业大学 | Sterile micro-cuttage rapid propagation method for populus deltoides |
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Application publication date: 20161012 |