CN104381132B - Rooting method of Juglans hindsii*J.regia vlach tissue culture seedling in bottle - Google Patents
Rooting method of Juglans hindsii*J.regia vlach tissue culture seedling in bottle Download PDFInfo
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Abstract
The invention discloses a rooting method of a Juglans hindsii*J.regia vlach tissue culture seedling in a bottle. A healthy Juglans hindsii*J.regia vlach tissue culture seedling in a subculture period is used as a rooting material and is treated by dark culture; then the Juglans hindsii*J.regia vlach tissue culture seedling is conveyed into a rooting culture medium to be subjected to rooting culture with alternative light culture and dark culture; a tissue culture rooting seedling which is planted in the bottle can be obtained after 20-24 days; and the rooting rate is up to 65%-75% and the greenhouse survival rate is up to 65%-85%. The rooting method of the Juglans hindsii*J.regia vlach tissue culture seedling in the bottle disclosed by the invention is not influenced by factors including seasons and the like, and theoretical basis and technical supports are provided for industrialized production, and uniform and stable germchits can be efficiently and rapidly provided; and the popularization speed of improved variety is accelerated. The rooting method is applicable to rooting culture of the Juglans hindsii*J.regia vlach tissue culture seedling in the bottle.
Description
Technical field
The invention belongs to domestication of plants field, relate to a kind of plant establishment method, be specifically related to a kind of Paradox walnut crow and draw
Rooting method in gram tissue cultured seedling bottle.
Background technology
Semen Juglandis is a kind of nonwood forest trees being difficult to and taking root, and there were significant differences for different IPs Peach cultivars rooting characteristics.According to the U.S.
University of California Gale McGranahan report in 2004, the holy cold spy of 3 Semen Juglandis main breeds that laboratory is used
(Sunlan), in strong Teller (Chandler), wiener (Vina), situation of taking root is widely different, and growth potential is strong, the product that tree performance is upright
Planting holy cold spy the most easily to take root, rooting rate reaches 94%；Strong Teller rootability is in middle level, reaches 55%；Wiener is then
The kind that difficulty is taken root, rooting rate only has 27%.This research also confirms that Walnut Cultivars or genotype are to determine that the raw test tube Seedling of Semen Juglandis is raw
The key factor of root.
" progress of Semen Juglandis rooting of vitro seedling " Li Chunyan etc., " north gardening ", public in page 2013 (01): 190-193
Having opened Walnut Cultivars different, genotype is different, and the expression of its rootability or potential of taking root there is also larger difference, shows not
With the difference of ability great disparity that adventitious root occurs between seeds and kind individuality.
" impact that Semen Juglandis tissue cultured seedling is taken root by IBA and dark treatment " Li Chunyan etc., " Shanxi Forestry science and technology ", 2011 12
Month, the 4th phase of volume 40,30-31 page, for examination material, inquire into IBA concentration and dark treatment sky with " rich fragrant No. 6 of gold " Semen Juglandis tissue cultured seedling
Several impacts taking root Semen Juglandis tissue cultured seedling, under its optimum condition, rooting rate is up to 65.56%.And point out: different auxin
Different to Semen Juglandis root induction effect.Adventitious root production process depends not only on the content of Endogenous auxin, also relates to hormone
Between balance.
" J．sigllata Dode isolated culture and Fast-propagation " Wang Shasha etc., " plant physiology journal ", 2014,50 (4), 527-537
Page, disclosing the stem-segment with single bud with fine individual plant " Nayong-1 " is outer implant, establishes J．sigllata Dode (Juglans sigillata)
Isolated culture and the system of Fast-propagation, its highest rooting rate is 71.73%,
And Paradox walnut (Juglans hindsii × J.regia) belongs to Juglandaceae (Juglandales) Juglans
(Juglans), being the cenospecies of California, U.S. north black walnut and Semen Juglandis, wherein male parent is Semen Juglandis, and female parent is California, north black walnut.
This seeds crow clarke Vlach is the elite plant strain selected from Paradox walnut, is a Semen Juglandis stock variety, in 2010 through state
Interior famous Semen Juglandis expert Xi Shengke professor introduces from California, USA, and it has obvious hybrid vigor, hence it is evident that be better than existing core
Peach rootstock, growth potential is good, has nematicide, Phytophthora pathogenic bacteria, root nodule bacteria, the disease that Armillaria mellea etc. causes.Bioclimatic analysis is wide, energy
Normal growth on the soil of pH ＞ 5, normal growth on the calcium soil of pH=9.This kind has certain winter resistance, suitable growth
In the abundant area of rain profit, the region of rainfall ＞ 500mm all can normal growth.Owing to its growing way is prosperous, it is particularly suitable as the most real core
The stock of Fructus Persicae, strengthens tree vigo(u)r.Doing stock with it, general 6 years i.e. up to the first best fruiting period.Meanwhile, these seeds are hard wealthy arbor of falling leaves,
Growth potential is strong, and trunk is logical straight, is urban greening and the excellent material of farmland woods greening, and compared with black walnut, its material is the most shallow, is situated between
Between black walnut and Semen Juglandis, can use as hard wealthy material, this type of timber many genus import the most domestic.
" research of Paradox walnut fast breeding technique " Qiu Xiaomei etc., " Inner Mongol forestry science and technology ", volume 39, the 2nd phase, 52-54 page,
Disclosing with Paradox walnut stem segment with axillary bud is that outer implant carries out Propagations of Teat Tube Seedlings, axillalry bud germinating culture medium and differentiation, successive transfer culture
It is DKW+IBA 1.0mg/L.Using " two step method " root induction effect preferable, the suitableeest root induction culture medium is DKW+IBA
2.5mg/L, root media is the DKW without phytohormone, and rooting rate can reach 65%.Owing to Paradox walnut belongs to cenospecies,
The Paradox walnut Seedling gone out of each seed culture belongs to different genotype.
And Paradox walnut is scion on domestic market, strain source confusion, there is no clone and take root Seedling.Singular kernel
Fructus Persicae (vlach) branch cutting rooting rate is low, and plant tissue culture, as vegetative manner, becomes and carried out by this superior clone
The inevitable mode of breeding, but due to the problem of its difficulty of taking root, the most not yet form industrialization.Nursery stock markets coker Peach rootstock at present
Various in style, very different, and mostly be planting seed and obtain, there is the problem of trait segregation in offspring, merit is difficult to stable
Heredity, though there being the report of Semen Juglandis tissue-culturing rapid propagation, but owing to its difficulty is taken root in existing research, it is impossible to forms factorial praluction.Therefore,
The rooting method of a kind of Paradox walnut (vlach) of research, and promoted and the economic benefit of forestry industry will be greatly improved, have
Important meaning.
Summary of the invention
The technical problem to be solved in the present invention, is to provide the rooting method in a kind of Paradox walnut crow clarke tissue cultured seedling bottle,
This rooting method, is not affected by factors such as seasons, provides theoretical foundation and technical support for its factorial praluction, and can be high
The seedling of effect, quickly offer stable homogeneous, accelerates improved variety popularization speed, and rooting rate can reach 65-75%, booth survival rate
Reach 65-85%.
For solving above-mentioned technical problem, the technical solution used in the present invention is:
A kind of rooting method in Paradox walnut crow clarke tissue cultured seedling bottle, it comprises the following steps:
(1) choose plant height be 2-5cm, subculture cycle be 18-23 days, leaf color be green or cyan Paradox walnut
Tissue cultured seedling reserves the stem section of 1-2 centimetre as material of taking root, excision radical leaves, axillalry bud to base portion, obtains material a of taking root；
(2) carrying out light culture in I culture medium of the seedling base portion taken root in material a insertion being taken root, the light culture time is 48-
72 hours, temperature was 20-28 DEG C, obtained material b of taking root；
(3) material b of taking root proceeds to take root in II culture medium, and alternately, illumination cultivation condition is for illumination and light culture
Illumination in 13 hours, intensity of illumination 2000-3000 lux, temperature is 23-27 DEG C；Light culture condition is light culture 11 hours, temperature
Degree, for 19-23 DEG C, obtains material c of taking root；Wherein, II culture medium of taking root be volume ratio be the nutritional solution of 1:1: substrate；
(4) material c of taking root cultivates 20-24 days, prepares Paradox walnut crow clarke group and trains Seedling of taking root.
As a kind of restriction of the present invention, I culture medium of taking root in described step (2) is: with the 1/4-1 special training of part Semen Juglandis
Supporting culture medium based on base DKW, add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-
6.0。
As the another kind of restriction of the present invention, the nutritional solution in described step (3) is with 1/4-1 part Semen Juglandis special culture media
Culture medium based on DKW, adds 0.01-0.03mg/L indolebutyric acid IBA and 20-50g/L sucrose, and pH is 5.5-6.0；
The present invention also has one to limit, and described substrate is Vermiculitum or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are
The Vermiculitum of 1:1:1: the peat composed of rotten mosses: the one in perlite.
Rooting efficiency corresponding to rooting method provided by the present invention is in view of all technology involved in rooting method
Effect after parameter combination, individually use therein one or more all can not reach corresponding technique effect, by therein
One or more technical parameters can not reach corresponding technique effect after simple replacement.
Owing to have employed above-mentioned technical scheme, compared with prior art, acquired technological progress is the present invention:
The present invention uses the rooting method in Paradox walnut crow clarke tissue cultured seedling bottle, is that plant tissue culture enters batch production
Produce an indispensable step.Wherein, using to the healthy Paradox walnut crow clarke tissue cultured seedling of subculture cycle as material of taking root, incite somebody to action
It processes through light culture, then proceeds to root media light culture and dark culture alternately root culture, after 20-24 days i.e.
The group that can obtain bottle outlet plantation trains Seedling of taking root, and its rooting rate reaches 65-75%, booth survival rate 65-85%.Provided by the present invention
Rooting method in Paradox walnut crow clarke tissue cultured seedling bottle, is not affected by factors such as seasons, provides reason for its factorial praluction
Opinion foundation and technical support, and the energy seedling providing stable homogeneous efficient, quick, accelerate improved variety popularization speed.
The present invention is applicable to the cultivation of taking root in Paradox walnut crow clarke tissue cultured seedling bottle.
The present invention is described in further detail below in conjunction with specific embodiment.
Detailed description of the invention
Rooting method in 1 one kinds of Paradox walnut crow clarke tissue cultured seedling bottles of embodiment
A kind of rooting method in Paradox walnut crow clarke tissue cultured seedling bottle, it comprises the following steps:
(1) choose plant height be 3cm, subculture cycle be 20 days, leaf color be the training of green or cyan Paradox walnut group
As taking root material, excision radical leaves, axillalry bud reserve 1 to base portion to Seedling (come from China forest-science academy Xi Shengke and teach laboratory)
Centimetre stem section, obtain material a of taking root；
(2) carrying out light culture in I culture medium of the seedling base portion taken root in material a insertion being taken root, the light culture time is 36
Hour, temperature is 24 DEG C, obtains material b of taking root；
Wherein, I culture medium of taking root is: culture medium based on 1 part of Semen Juglandis special culture media DKW, adds 8mg/L indole fourth
Acid IBA, 50g/L sucrose and 6g/L agar, pH is 5.5.
(3) material b of taking root proceeds to take root in II culture medium, and alternately, illumination cultivation condition is for illumination and light culture
Illumination in 13 hours, intensity of illumination 2500 lux, temperature is 25 DEG C；Light culture condition is light culture 11 hours, and temperature is 21 DEG C,
Obtain material c of taking root；Wherein, II culture medium of taking root be volume ratio be the nutritional solution of 1:1: substrate；
Wherein, nutritional solution is culture medium based on 1/4-1 part Semen Juglandis special culture media DKW, adds 0.01mg/L indole
Butanoic acid IBA and 30g/L sucrose, pH is 5.5；
Described substrate is Vermiculitum or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are the Vermiculitum of 1:1:1: the peat composed of rotten mosses: precious
One in pearl rock.
(4) material c of taking root cultivates 24 days, prepares Paradox walnut crow clarke group and trains Seedling of taking root, and rooting rate is 75%, booth
Rate of growth is 85%.
Rooting method in embodiment 2-6 Paradox walnut crow clarke tissue cultured seedling bottle
Embodiment 2-6 is respectively the rooting method in a kind of Paradox walnut crow clarke tissue cultured seedling bottle, itself and embodiment 1 phase
Seemingly, the difference is that only that technical parameter involved in method is different, referring specifically to following table:
Wherein, take root I culture medium and the raw material components taken root in II culture medium and the consumption such as following table institute in embodiment 2-6
Show:
Embodiment 7 is taken root the screening of condition
One, the screening of the dark treatment temperature and time in step (2)
| Experimental group | Dark treatment temperature (DEG C) | Dark treatment time (h) | Rooting rate (%) | Booth survival rate (%) |
| 1 | 15 | 36 | 15 | 8 |
| 2 | 18 | 42 | 23 | 40 |
| 3 | 18 | 48 | 62 | 53 |
| 4 | 20 | 42 | 50 | 67 |
| 5 | 20 | 48 | 72 | 83 |
| 6 | 23 | 54 | 70 | 80 |
| 7 | 25 | 63 | 74 | 85 |
| 8 | 28 | 72 | 75 | 81 |
| 9 | 30 | 78 | 68 | 55 |
| 10 | 28 | 78 | 70 | 58 |
| 11 | 25 | 72 | 63 | 72 |
As seen from the above table, optimal dark treatment temperature is 20-28 DEG C, and the dark treatment time is 48-72h, rooting rate and booth
Survival rate, respectively reaches 70-75% and 80-85%；But when dark treatment temperature and time is all not up to range lower limit or exceeds
During range limit, rooting rate and booth survival rate are relatively low；Temperature therein in the range of but the time go beyond the scope or the time exists
In the range of but temperature is when going beyond the scope, rooting rate and booth survival rate be also significantly lower than in the range of result.Reason is dark place
Reason overlong time can cause plant yellow, aging occur, fall leaves and produce callus etc., and adventitious root is produced harmful effect.
Two, I culture medium of taking root and the screening of raw material components consumption taken root in II culture medium
Basal medium uses DKW culture medium, is added to IBA, sucrose and agar, and the pH value of regulation is for life
Root effect plays very important effect.The amount of the minimum use of DKW culture medium is 1/4 part, if usage amount reduces again, just
Necessary nutrient substance can not be provided to the seedlings cultivating needing to take root, if 1 part, its rooting rate and booth can not be made raw
Long rate is significantly improved, and also can cause the increase of production cost, and therefore the consumption of the DKW culture medium of final choice is 1/4-
1 part.
As seen from the above table, the optimum proportioning of I culture medium of taking root is for training based on 1/4-1 part Semen Juglandis special culture media DKW
Supporting base, add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-6.0.Beyond being provided
Lower limit rooting rate and the booth survival rate of scope are decreased obviously, because its nutritional labeling provided fails to reach seedlings cultivating growth
Required requirement；Higher limit rooting rate and booth survival rate beyond provided scope the most significantly increase, and can lead
Cause the increase of production cost.
As seen from the above table, the optimum proportioning of II culture medium of taking root is for training based on 1/4-1 part Semen Juglandis special culture media DKW
Supporting base, add 0-0.03mg/L indolebutyric acid IBA and 20-50g/L sucrose, pH is 5.5-6.0；Lower limit beyond provided scope
Value rooting rate and booth survival rate are decreased obviously, because its nutritional labeling provided fails to reach wanting needed for seedlings cultivating grows
Ask；Higher limit rooting rate and booth survival rate beyond provided scope the most significantly increase, and can cause producing into
This increase.
By the screening to above Paradox walnut crow clarke tissue cultured seedling root media, finishing screen is selected suitable its and is taken root
Culture medium prescription and concrete rooting method, the highest rooting rate reaches 75%, and root system is the normal root system of white, through booth
After seedling exercising, booth survival rate is up to 85%.Efficiently solve Paradox walnut crow clarke tissue cultured seedling take root difficulty problem, for its work
Factory's metaplasia is produced provides theoretical foundation and technical support, and this technology is not by seasonal effect, can produce stable homogeneous throughout the year
The training of Paradox walnut crow clarke group take root Seedling.After it carries out factorial praluction, the most only nursery stock markets provides stabilization characteristics of genetics
Paradox walnut crow clarke stock seeds, it is possible to provide substantial amounts of Timber stands as reproducting tree species, finally bring for market huge
Economic benefit and ecological benefits.
The comparison of embodiment 8 rooting method
Compared for " one-step method is taken root ", " two-step method is taken root " and rooting method provided by the present invention, result is as follows:
" one-step method is taken root " of the prior art refers to that test tube Seedling is being added with in the culture medium of auxin cultivation always.
" two-step method is taken root " refers to test tube Seedling root induction certain time in a kind of culture medium, then proceeds to without auxin
Culture medium continues cultivate.
Owing to Paradox walnut is taken root more difficult, use one-step method to be difficult to its phenomenon of taking root, and Semen Juglandis is to auxin
Class material is the most sensitive, its basal part of stem uses the auxin of higher concentration process for a long time, can make its enlargement and produce
Raw callus, this root system produced at callus is not communicated with the conducting system of stem, is unfavorable for the growth of sprout, very
To root can be made to come off；If reducing auxin concentration used, occur almost without adventitious root again.
| Rooting method | Rooting rate (%) | Booth rate of growth (%) |
| " two-step method " | 25.3-65 | 83 |
| The rooting method of the present invention | 65-75 | 65-85 |
As seen from the above table, using Paradox walnut crow clarke tissue cultured seedling as material of taking root, provided by the present invention taking root is used
Rooting rate and booth rate of growth obtained by method are the highest.Also the tissue cultured seedling of explanation Paradox walnut crow clarke kind is the most uncomfortable
For the method that prior art is general.
The above, be only presently preferred embodiments of the present invention, is not the restriction that the present invention makees other form, appoints
What those skilled in the art is changed possibly also with above-mentioned technology contents as enlightenment or is modified as equivalent variations
Equivalent embodiments.But every without departing from technical solution of the present invention content, the technical spirit of the foundation present invention is to above example
Done simple modification, equivalent variations and remodeling, still fall within the protection domain of the claims in the present invention.
Claims (2)
1. the rooting method in a Paradox walnut crow clarke tissue cultured seedling bottle, it is characterised in that it comprises the following steps:
(1) choose plant height be 2-5cm, subculture cycle be 18-23 days, leaf color be the training of green or cyan Paradox walnut group
Seedling reserves the stem section of 1-2 centimetre as material of taking root, excision radical leaves, axillalry bud to base portion, obtains material a of taking root；
(2) carrying out light culture in I culture medium of the seedling base portion insertion of material a of taking root being taken root, the light culture time is that 48-72 is little
Time, temperature is 20-28 DEG C, obtains material b of taking root；Described I culture medium of taking root is: with 1/4-1 part Semen Juglandis special culture media DKW
Based on culture medium, add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-6.0；
(3) material b of taking root proceeds to take root in II culture medium, and alternately, illumination cultivation condition is 13 little for illumination and light culture
Shi Guangzhao, intensity of illumination 2000-3000 lux, temperature is 23-27 DEG C；Light culture condition is light culture 11 hours, and temperature is
19-23 DEG C, obtain material c of taking root；Wherein, II culture medium of taking root be volume ratio be the nutritional solution of 1:1: substrate；Described nutritional solution
For culture medium based on 1/4-1 part Semen Juglandis special culture media DKW, add 0.01-0.03mg/L indolebutyric acid IBA and 20-
50g/L sucrose, pH is 5.5-6.0；
(4) material c of taking root cultivates 20-24 days, prepares Paradox walnut crow clarke group and trains Seedling of taking root.
Rooting method in Paradox walnut crow clarke tissue cultured seedling bottle the most according to claim 1, it is characterised in that: described base
Matter is Vermiculitum or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are the Vermiculitum of 1:1:1: the peat composed of rotten mosses: the one in perlite.
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| CN110741934A (en) * | 2019-11-22 | 2020-02-04 | 塔里木大学 | Method for rooting in early-fruiting walnut tissue culture seedling bottle |
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