CN108308023A - A kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method - Google Patents
A kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method Download PDFInfo
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- CN108308023A CN108308023A CN201810037076.XA CN201810037076A CN108308023A CN 108308023 A CN108308023 A CN 108308023A CN 201810037076 A CN201810037076 A CN 201810037076A CN 108308023 A CN108308023 A CN 108308023A
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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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- 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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Abstract
The invention belongs to technical field of plant culture, and in particular to a kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method.Include the selection and inoculation of saline-alkali tolerant fast-growing white elm clones adventitious bud, the condition of adventitious bud rooting is：(EM take root+IBA1mg/L+2%S) is carried out in the formula researched and developed of the present invention, cultivation temperature is 18 DEG C of (± 2 DEG C)/night of 26 DEG C of daytime (± 2 DEG C), 90% environment culture of relative air humidity, daily illumination 12 hours, 2000-3500 lux of intensity of illumination.Formula using the present invention and method, American elm adventitious bud rooting rate is up to 96%；And the adventitious bud stock utilization for taking root improves 20 35% or more than early period.
Description
Technical field
The invention belongs to technical field of plant culture, and in particular to a kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture
Rooting method.
Technical background
American elm (UlmuspumilaL.), is Ulmaceae (Ulmaceae) Elm (Ulmus) plant, and mutation is more.
Salt-soda soil be salt in the soil inside aggregation and soil contained by salinity influenced the one of plant normal growth
Kind land type.The saline alkali land area in China is 3.5 × 107hm2, it is equivalent to the 33.3% of cultivated area.How salt is efficiently used
The ecological environment in alkali area improves the xylophyta coverage rate in salt-soda soil, is one of the important problem that forest workers is faced.
American elm is as one of main gardens native soil ornamental tree species in China, to the toxic gases resistance such as flue dust, sulfur dioxide and hydrogen fluoride
By force, it and with high environmental suitability, antipollution, Resistant, preferably breeds and stronger saline-alkali tolerant characteristic so that American elm pole
Promise to be varieties in saline-alkali areas afforestation Key species.
In order to ensure that descendant inheritting stability, saline-alkali tolerant fast-growing white elm clones mainly pass through cuttage, grafting and tissue
The asexual manners such as culture are bred, wherein cuttage, engrafting method, that is, time-consuming and laborious, and are difficult to ensure survival rate.Therefore, sharp
With the mode of Plant Tissue Breeding, carries out saline-alkali tolerant fast-growing white elm clones and a kind of actually quickly and effectively side is factory produced
Method.
It is studied for rooting method in saline-alkali tolerant fast-growing white elm clones adventitious bud bottle, in 1/2WPM+ early period
Under sugared 20% condition of culture of IBA0.2mg/L+NAA0.1mg/L+ activated carbons 0.5g/L+ in clone bottle rooting rate up to 95%,
Substantially meet factorial praluction demand.But since the white elm clones adventitious bud for taking root in bottle has strict requirements
(height of seedling must be 2-3.5cm, the individual that healthy and strong, blade is unfolded), causes greatly to waste in reality produces.Activated carbon is being cultivated
Base easily precipitates when solidifying, and unnecessary trouble is manufactured into matching for root media.Therefore, for saline-alkali tolerant fast-growing American elm
Very big Development volue, it would be highly desirable to develop rooting method in a kind of simple, the efficient adventitious bud bottle of utilization.
Invention content
It is an object of the invention to provide one for saline-alkali tolerant fast-growing white elm clones tissue-cultured seedling suitable for raw in industrial bottle
Method for root.The efficiency of saline-alkali tolerant fast-growing American elm factorial praluction is improved using rooting method in this bottle, improves stock utilization,
Simplified culture base preparation method reduces production cost, increases economic efficiency.
To achieve the above object, the invention is realized by the following technical scheme：
A kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method, used medium of taking root are EM culture of rootage
Base, including following components and its content：CaCl2·2H2O 200-240mg/L, NH4NO3600-1000mg/L, KNO3 1000-
1200mg/L, MgSO41400-1800mg/L, NaH2PO4·2H2O 100-200mg/L, K2SO4100-250mg/L, Na2SO4
100-200mg/L, CaCl2100-130mg/L, CoCl2·6H2O 0.15-0.4mg/L, CuSO4·5H2O 0.325-
0.415mg/L, H3BO38-10mg/L, MnSO4·H2O 15-17mg/L, Na2MoO4·2H2O 0.1-0.4mg/L, ZnSO4·
7H2O 10-13mg/L, KI 0.6-0.9mg/L, FeSO4·7H2O 26-29mg/L, Na2EDTA·2H2O 36-38mg/L, flesh
Alcohol 100-120mg/L, glycine 2-4mg/L, niacin 4-6mg/L, puridoxine hydrochloride 1.0-1.4mg/L, Tyiamine Hd element 12-
14mg/L, biotin 0.1-0.3mg/L, folic acid 0.7-0.9mg/L, vitamin C 1.52-1.85mg/L, riboflavin 1.5-
2.2mg/L。
Further, used medium of taking root is that EM root medias include following components and its content：CaCl2·2H2O
220.5mg/L, NH4NO3800mg/L, KNO31011mg/L, MgSO41680.5mg/L NaH2PO4·2H2O 156mg/L,
K2SO4175mg/L, Na2SO4150mg/L, CaCl2115mg/L, CoCl2·6H2O 0.24mg/L, CuSO4·5H2O
0.375mg/L, H3BO39.3mg/L, MnSO4·H2O 16.9mg/L, Na2MoO4·2H2O 0.25mg/L, ZnSO4·7H2O
11.5mg/L, KI 0.83mg/L, FeSO4·7H2O 27.8mg/L, Na2EDTA·2H2O 37.25mg/L, inositol 108mg/L,
Glycine 3.75mg/L, niacin 5mg/L, puridoxine hydrochloride 1.2mg/L, Tyiamine Hd element 13.5mg/L, biotin 0.2mg/L,
Folic acid 0.88mg/L, vitamin C 1.76mg/L, riboflavin 1.9mg/L.
Further, the EM root medias further include IBA1mg/L.
Further, the EM root medias further include sucrose 20g/L.
Further, the EM root medias of stating further include agar 6.5g/L.
Further, the pH value of the EM root medias is 5.8-6.0.
Culture of rootage is carried out to saline-alkali tolerant fast-growing white elm clones adventitious bud with EM root medias of the present invention, rooting rate is high
Reach 96%.Compared with EM culture mediums, CaCl in EM root medias of the present invention2·2H2O、NH4NO3、KNO3、MgSO4、
NaH2PO4·2H2O、K2SO4、Na2SO4、CaCl2Ingredient Amount reduces half, and is free of activated carbon, and EM of the present invention is taken root training
Foster base greatly reduces the dosage of raw material, increases the utilization rate of raw material, i.e., is significantly reduced during being factory produced
Production cost；Each ingredient interaction, has effectively facilitated saline-alkali tolerant fast-growing white elm clones not in EM root medias of the present invention
Normal bud takes root.
Further, saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method the specific steps are：
(1) selection taken root with adventitious bud：Growth selection is normal, through adopt with culture and/or callus be differentiated to form it is resistance to
Saline and alkaline fast-growing white elm clones adventitious bud；
(2) adventitious bud is inoculated with：Qualified saline-alkali tolerant white elm clones adventitious bud is inoculated in EM root medias, is connect
Kind is placed on 18 ± 2 DEG C of 26 ± 2 DEG C/night of daytime, 90% environment culture of relative air humidity, and carries out photo-irradiation treatment.
Further, adventitious bud plant height 1.5cm or more is selected in step (1).It is same using EM root medias culture of the present invention
Sample reduces the requirement for using taking root adventitious bud, adventitious bud plant height 1.5cm or more.
Further, 2000-3500 lux of intensity of illumination in step (2).
Further, daily illumination 12 hours in step (2).
Compared with prior art, the present invention has the following advantages：
(1) present invention uses EM root medias, does not add activated carbon, and reduce the use of Multiple components in EM culture mediums
Amount, rooting rate can reach 96%, and the present invention greatly reduces the dosage of raw material, increases the utilization rate of raw material, i.e., in factory
Production cost is significantly reduced during metaplasia production；
(2) present invention reduces using EM root medias are used and requires plant height 1.5cm or more i.e. to the screening of adventitious bud
Can, individual growth is neat after taking root, and growing way is vigorous, and the present invention greatly improves production efficiency, increases economic benefit.
Description of the drawings
Fig. 1 is：American elm adventitious bud rooting situation after a week；
Fig. 2 is：After 25 days, American elm adventitious bud rooting situation before transplanting, left figure is adventitious root close up fragmentary, and right figure is American elm
Adventitious bud rooting situation；
Fig. 3 is：It takes root transplantation of seedlings growing state after a week.
Specific implementation mode
It is noted that described further below be all exemplary, it is intended to provide further instruction to the present invention.Unless another
It indicates, all technical and scientific terms used herein has usual with general technical staff of the technical field of the invention
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific implementation mode, and be not intended to restricted root
According to exemplary embodiments of the present invention.As used herein, unless the context clearly indicates otherwise, otherwise singulative
It is also intended to include plural form, additionally, it should be understood that, when in the present specification using term "comprising" and/or " packet
Include " when, indicate existing characteristics, step, operation, component and/or combination thereof.
In order to enable those skilled in the art can clearly understand technical scheme of the present invention, below with reference to tool
The embodiment of the body technical solution that the present invention will be described in detail.
Embodiment
(1) selection taken root with adventitious bud：Growth selection is normal, plant height 1.5cm or more, by repeatedly after with culture (or
Callus is differentiated to form) saline-alkali tolerant fast-growing white elm clones adventitious bud (growth cycle is less than except one week tender tissue)；
(2) adventitious bud is inoculated with：Qualified saline-alkali tolerant white elm clones adventitious bud is inoculated in EM root medias, into
Row culture of rootage.
Growing environment is：18 DEG C of (± 2 DEG C)/night of 26 DEG C of daytime (± 2 DEG C), daily illumination 12 hours, intensity of illumination
2000-3500 luxs, relative air humidity 90%.
EM root medias include following components and its content：CaCl2·2H2O 220.5mg/L, NH4NO3800mg/L,
KNO31011mg/L, MgSO41680.5mg/L NaH2PO4·2H2O 156mg/L, K2SO4175mg/L, Na2SO4 150mg/
L, CaCl2115mg/L, CoCl2·6H2O 0.24mg/L, CuSO4·5H2O 0.375mg/L, H3BO39.3mg/L, MnSO4·
H2O 16.9mg/L, Na2MoO4·2H2O 0.25mg/L, ZnSO4·7H2O 11.5mg/L, KI 0.83mg/L, FeSO4·7H2O
27.8mg/L Na2EDTA·2H2O 37.25mg/L, inositol 108mg/L, glycine 3.75mg/L, niacin 5mg/L, hydrochloric acid pyrrole are trembled
Alcohol 1.2mg/L, Tyiamine Hd element 13.5mg/L, biotin 0.2mg/L, folic acid 0.88mg/L, vitamin C 1.76mg/L, core yellow
Plain 1.9mg/L, sucrose 20g/L, agar 6.5g/L, IBA1mg/L.
Culture medium preparation condition：Experiment adjusts Medium's PH Value to 5.9 (before sterilizings) with the NaOH and HCl of 1mol/L, and
In 121 DEG C, 1.1Kg/cm2(0.11MPa) sterilizing 14min.
Test example
Rooting method of the present invention is carried out with the method taken root in the prior art using the culture medium containing activated carbon
Compare, is grouped as follows：
M1 groups：1/2WPM+IBA1mg/L+ sucrose 20%；
M2 groups：1/2WPM+IBA0.2mg/L+NAA0.1mg/L+ activated carbon 0.5g/L+ sucrose 20%
M3 groups：EM root medias of the present invention, are cultivated by above-described embodiment method.
Experiment process result：
By 25 days experiment process, in each culture medium under the conditions of white elm clones adventitious bud rooting situation, rooting rate statistics
Refer to table 1.
1 white elm clones adventitious bud rooting situation statistical form of table
As can be seen from Table 1, saline-alkali tolerant fast-growing white elm clones adventitious bud rooting rate in the case where the present invention cultivates treatment conditions
Up to 96%, rooting rate improves 1% compared with M3 groups, illustrates that activated carbon is not added in culture of rootage of the present invention can also effectively facilitate life
Root, and improve 20-35% or more for method before the adventitious bud stock utilization ratio of rooting treatment.M3 groups of the present invention 14 days
Rooting rate is 2 times of M1 groups, and rooting rate improves 14% compared with M1 groups within 25 days；It can thus be appreciated that each ingredient of EM culture mediums of the present invention
Between have synergistic effect.Rooting method of the present invention reduces production cost under the premise of improving production American elm production quantity,
In large-scale production, higher economic benefit is brought.
The above shows and describes the basic principles and main features of the present invention and the advantages of the present invention.The technology of the industry
Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this
The principle of invention, without departing from the spirit and scope of the present invention, various changes and improvements may be made to the invention, these changes
Change and improvement all fall within the protetion scope of the claimed invention, the claimed scope of the invention by appended claims and its
Equivalent thereof.
Claims (10)
1. a kind of saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture rooting method, characterized in that used medium of taking root is EM
Root media includes following components and its content：CaCl2·2H2O 200-240mg/L, NH4NO3600-1000mg/L, KNO3
1000-1200mg/L, MgSO41400-1800mg/L, NaH2PO4·2H2O 100-200mg/L, K2SO4100-250mg/L,
Na2SO4100-200mg/L, CaCl2100-130mg/L, CoCl2·6H2O 0.15-0.4mg/L, CuSO4·5H2O
0.325-0.415mg/L, H3BO38-10mg/L, MnSO4·H2O 15-17mg/L, Na2MoO4·2H2O 0.1-0.4mg/L,
ZnSO4·7H2O 10-13mg/L, KI 0.6-0.9mg/L, FeSO4·7H2O 26-29mg/L, Na2EDTA·2H2O 36-
38mg/L, inositol 100-120mg/L, glycine 2-4mg/L, niacin 4-6mg/L, puridoxine hydrochloride 1.0-1.4mg/L, hydrochloric acid sulphur
Ammonium element 12-14mg/L, biotin 0.1-0.3mg/L, folic acid 0.7-0.9mg/L, vitamin C 1.52-1.85mg/L, riboflavin
1.5-2.2mg/L。
2. method according to claim 1, which is characterized in that used medium of taking root is that EM root medias include with the following group
Point and its content：CaCl2·2H2O 220.5mg/L, NH4NO3800mg/L, KNO31011mg/L, MgSO41680.5mg/L
NaH2PO4·2H2O 156mg/L, K2SO4175mg/L, Na2SO4150mg/L, CaCl2115mg/L, CoCl2·6H2O
0.24mg/L, CuSO4·5H2O 0.375mg/L, H3BO39.3mg/L, MnSO4·H2O 16.9mg/L, Na2MoO4·2H2O
0.25mg/L, ZnSO4·7H2O 11.5mg/L, KI 0.83mg/L, FeSO4·7H2O 27.8mg/L, Na2EDTA·2H2O
37.25mg/L, inositol 108mg/L, glycine 3.75mg/L, niacin 5mg/L, puridoxine hydrochloride 1.2mg/L, Tyiamine Hd element
13.5mg/L, biotin 0.2mg/L, folic acid 0.88mg/L, vitamin C 1.76mg/L, riboflavin 1.9mg/L.
3. method according to claim 1, which is characterized in that above-mentioned EM root medias further include IBA1mg/L.
4. method according to claim 1, which is characterized in that above-mentioned EM root medias further include sucrose 20g/L.
5. method according to claim 1, which is characterized in that above-mentioned EM root medias further include agar 6.5g/L.
6. method according to claim 1, which is characterized in that the pH value of the EM root medias is 5.8-6.0.
7. according to any the methods of claim 1-6, which is characterized in that saline-alkali tolerant fast-growing white elm clones adventitious bud tissue culture is given birth to
Method for root the specific steps are：
(1) selection taken root with adventitious bud：Growth selection is normal, through the saline-alkali tolerant that with is cultivated and/or callus is differentiated to form of adopting
Fast-growing white elm clones adventitious bud；
(2) adventitious bud is inoculated with：Qualified saline-alkali tolerant white elm clones adventitious bud is inoculated in EM root medias, after inoculation
It is placed in 18 ± 2 DEG C of 26 ± 2 DEG C/night of daytime, 90% environment culture of relative air humidity, and carries out photo-irradiation treatment.
8. method according to claim 7, which is characterized in that select adventitious bud plant height 1.5cm or more in step (1).
9. method according to claim 7, which is characterized in that 2000-3500 lux of intensity of illumination in step (2).
10. method according to claim 7, which is characterized in that daily illumination 12 hours in step (2).
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