CN108651274A - Diversiform-leaved poplar seed for salt stress processing60Co- gamma Rays method of mutagenesis - Google Patents
Diversiform-leaved poplar seed for salt stress processing60Co- gamma Rays method of mutagenesis Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
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Abstract
The invention discloses the diversiform-leaved poplar seeds handled for salt stress60Co gamma Rays method of mutagenesis, be by diversiform-leaved poplar seed with60After Co gamma-rays carries out radiation treatment, then the processing of diversiform-leaved poplar seed salt stress is carried out using sodium chloride solution and is cultivated.Beneficial effects of the present invention are：After being operated according to radioinduction method provided by the invention, germination percentage, germinating energy and the relative germination rate of 150Gy dose mutagenesis are significantly higher than other radiation treatments, and opposite salt damage rate reduces.By establishing diversiform-leaved poplar seed60Co gamma Rays mutation breeding technologies, foundation is provided for the selection and determination of the best dose of radiation of its radiaction mutation, disclose the sprouting mechanism that diversiform-leaved poplar seed coerces radioinduction and NaCl, the suitable dose of radiation of seed salt tolerance is provided, further provides new method and approach by radiaction mutation for the innovation of diversiform-leaved poplar germplasm resource and breed breeding.
Description
Technical field
The present invention relates to radioactive breeding technical fields, and in particular to is used for the diversiform-leaved poplar seed of salt stress processing60Co- γ are penetrated
Beta radiation method of mutagenesis.
Background technology
Radioinduction (physical mutagenesis) breeding is to carry out breeding using seed radiation technology, is that current breeding efficiency is higher
Technology mainly uses the radiation treatment material of doses, the mutant of inducing materials gene or being distorted for chromosome, warp
Screening, measurement, selection are crossed, to acquisition mutant in the short time, therefrom selects the process of new varieties.
Radioinduction is the ray using doses, and irradiation plant, pollen, seed or organ induce and become
It is different, it is allowed to generate the process being favorably mutated, new varieties is bred as by artificial screening and cultivation by.The premise of radioinduction is radiation
Processing, effect raying dosage, processing method, the influence for handling the factors such as material and region.Since people often ignore
Control to radiation condition can even cause phenomena such as irradiation is invalid or large area is lethal when serious.
In recent years, in radioinduction research, most common radioinduction source includes：Gamma-rays, X-ray, ultraviolet light, in
Son, laser, electron beam, ion beam etc..The gamma-ray radiation source that is usually widely used in plant breeding is60Co is being radiated
There is radiation condition to be easy to control, significant effect in terms of breeding.In addition to this, gamma-rays is that neutral ray, penetration power be relatively strong, spoke
It penetrates dose uniformity, can once handle a large amount of material, be one of first choice of radioactive breeding.Currently, radiaction mutation is because of tool
Have the advantages that mutation rate high, mutation spectrum width, fast, breeding cycle is short, enhancing resistance compared with stablizing keeping kind characters of progenies,
One as selection and breeding New idioplasm resource modernizes breeding technique, and continuously attempts to new skill in radioinduction field both at home and abroad
Art, method etc., and achieve prodigious achievement.
Diversiform-leaved poplar (Populus euphratica) is Salicaceae (Salicaceae) Populus (Populus L.) diversiform-leaved poplar subgenus
Plant is a kind of rare, ancient, endangered species, is grown on pole desert riverbank and in the unique of arid-desert area
Advantage is grown into forest seeds.Have the characteristics that very strong adaptability, saline-alkali tolerant, water is wet, drought-resistant dust storm, Populus Euphratica is distributed mainly on newly
Boundary Tarim Basin and the Junggar Basin, area account for the 91.1% of national Populus Euphratica area, and in Xinjiang, distribution is more concentrated, and is accounted for
90% or more of China's Populus Euphratica gross area.Diversiform-leaved poplar safeguards waste Mo River as the up to deciduous tree species in the riparian forest of oasis
The key species tree of the bank woods ecological balance contains desertification, checks winds and fixes drifting sand, regulate and control arid desert for improving region environment
The regional fragile ecosystem and protection bio-diversity etc. are of great significance.In recent years, due to large-scale water and soil
It develops (reclamations of wasteland), Cut off and the mankind cut excessively, denudate etc. artificially and the influence of natural cause, and Populus Euphratica is caused to face increasingly
It decays and situation in imminent danger, prestige is caused to diversiform-leaved poplar germ plasm resource since diversiform-leaved poplar deteriorates the importance of arid-desert area and habitat
The side of body.Therefore, imperative to diversiform-leaved poplar Germ-plasma resources protection.Recently, people have carried out many researchs to the breeding of diversiform-leaved poplar, but right
Diversiform-leaved poplar seed radiation breeding work is very few.
Mutation breeding has become one of main breeding technique, and new thinking is provided for plant breeding, is shown
Wide foreground.Radiaction mutation is applied in China's other plant breeding, but is answered in the plants such as diversiform-leaved poplar seed
With few.And seldom scholar by NaCl stress with60Diversiform-leaved poplar seed is studied in the processing of Co- gamma Rays.
Invention content
The purpose of the present invention aiming at above-mentioned defect in the prior art, provide a kind of germination percentage is high, salt tolerance is strong,
Save the diversiform-leaved poplar seed for salt stress processing of time cost60Co- gamma Rays method of mutagenesis.
The present invention uses various dose by material of diversiform-leaved poplar seed60Co- gamma Rays are handled, and in various concentration
NaCl under the conditions of carry out Seed Germination Test, explore60Co- gamma Rays coerce lower diversiform-leaved poplar Germination Characteristics to NaCl
It influences, can significantly promote diversiform-leaved poplar seed to sprout under NaCl stress to seek60Co- gamma Rays dosage, to spoke
It penetrates mutation breeding and provides new method and approach for diversiform-leaved poplar Germplasm enhancement and breed breeding.
To achieve the goals above, technical solution provided by the invention is：Diversiform-leaved poplar seed for salt stress processing60Co- gamma Rays method of mutagenesis, includes the following steps：
1) condition of culture is determined：
The processing of diversiform-leaved poplar seed salt stress, concentration of sodium chloride solution 0-300mmolL are carried out using sodium chloride solution-1, recklessly
The salt stress condition of culture of poplar seed is 23-27 DEG C of cultivation temperature, and intensity of illumination is 3 000lx, light dark period 16/8h；
2) radiation treatment：
By diversiform-leaved poplar seed with60Co- gamma-rays carries out radiation treatment,60Co- gamma Rays dosage is 0-200Gy, radiation
Time is 0-16min；Seed after radiation with 0.1% HgCl2Carry out disinfection 5~7min, with distilled water flushing 3 times, every time
1min takes sodium chloride solution 5ml to pour into the culture dish of paving two layers of filter paper, by 20 according still further to the concentration of sodium chloride solution of step 1)
Grain seed is inoculated in culture dish, is repeated 4 times, is sealed and be placed in constant temperature illumination box with sealed membrane, provided using step 1)
Condition of culture cultivated；In incubation, first water is filled daily to ensure each salinity in Seed Germination
It keeps constant, every the sprouting situation for counting seed for 24 hours, starts counting up within second day after inoculation, count 7d altogether.
Further, the above-mentioned diversiform-leaved poplar seed for salt stress processing60Co- gamma Rays method of mutagenesis, it is described
In salt stress processing in step 1), concentration of sodium chloride solution 181mmolL-1, cultivation temperature is 25-27 DEG C.
Further, the above-mentioned diversiform-leaved poplar seed for salt stress processing60Co- gamma Rays method of mutagenesis, it is described
In step 2), in the step 2),60Co- gamma Rays dosage is 150Gy, radiated time 0-16min.
Beneficial effects of the present invention are：Provided by the present invention for the diversiform-leaved poplar seed of salt stress processing60Co- gamma-rays spokes
Method of mutagenesis is penetrated, through various dose60The mutagenesis of Co- gamma Rays coerces lower progress Seed Germination Test to NaCl concentration, passes through
Variance analysis and Multiple range test analyze germination percentage and each germination index value.In difference60Co- gamma Rays mutagenesis pair
Under NaCl concentration stress, germination percentage, germinating energy and the relative germination rate of 150Gy dose mutagenesis are significantly higher than at other radiation
Reason, opposite salt damage rate reduce.The mutagenesis of 150Gy doses has certain facilitation, 181mmol to the sprouting of diversiform-leaved poplar seed
L-1It is a concentration of60Co- gamma Rays coerce NaCl the salt tolerant critical concentration value that lower diversiform-leaved poplar seed is sprouted, i.e., coerce NaCl
Show preferable tolerance.By establishing diversiform-leaved poplar seed60Co- gamma Rays mutation breeding technologies are its radioinduction
The selection of the best dose of radiation of breeding and determining offer foundation, disclose the sprouting that diversiform-leaved poplar seed coerces radioinduction and NaCl
Mechanism provides the suitable dose of radiation of seed salt tolerance, is further innovated for diversiform-leaved poplar germplasm resource by radiaction mutation
New method and approach are provided with breed breeding.
Description of the drawings
Fig. 1 is shown as60Difference of the Co- gamma Rays between the average germination percentage of diversiform-leaved poplar seed, germinating energy and germination index
It is different.
Fig. 2 is shown as60Co- gamma Rays sprout diversiform-leaved poplar seed the influence of stage salt tolerance.
Fig. 3 is shown as diversiform-leaved poplar seed items germination index average and exists60Difference of the Co- gamma Rays between NaCl stress.
Specific implementation mode
Embodiment 1：
Diversiform-leaved poplar seed provided by the invention60Co- gamma Rays method of mutagenesis, specific implementation mode are：
Select diversiform-leaved poplar seed, including step：
A, condition of culture：
The processing of seed salt stress：Salinity is set as NaCl solution, concentration be disposed as 0 (CK), 25,50,150,250,
300mmol/L, totally 6 Salt Strees Condition processing, wherein 1 control treatment.
The above condition of culture is 25-27 DEG C of temperature, intensity of illumination is 3 000lx；Light dark period is 16/8h.
B, radiation treatment：
Diversiform-leaved poplar seed is test material, and grouping is used60Co- gamma-rays carries out radiation treatment；
60Co- gamma Rays dosage (0,50,100,150,200Gy), seed after radiation with 0.1% HgCl2It carries out
5~7min is sterilized, with 3 each 1min of distilled water flushing.Experiment sets 0 (CK, distilled water), 25,50,150,250 and altogether
300mmol·L-1NaCl concentration gradient, is taken 5mL processing solutions to pour into the culture dish of paving two layers of filter paper, and every 20 seeds connect
Kind is in culture dish, and totally 6 processing, every group of processing is repeated 4 times, is sealed and be placed in constant temperature illumination box with sealed membrane, temperature
It is cultivated under conditions of for 25-27 DEG C, intensity of illumination be 3 000lx, light dark cycles are 16/8h.Supplement single flash daily
Water is to ensure that each salinity is kept constant in Seed Germination.Every the sprouting situation for counting seed for 24 hours.(after inoculation
Start counting up within two days) statistics 7d.Determine the salt tolerant critical concentration that the seed of radioinduction germinates under salt stress.
C, the germination index value of statistics seed, opposite salt damage rate and accumulation germination percentage：
Observation counts time and the chitting piece number of each dosage processing germination：Calculate germination index value with respect to salt damage rate and
Survival rate of seedling.
Germinate index determining：
Germination percentage (%)=(n/N) × 100%；N is the seed number that 7d normally germinates in formula, and N is for planting experimentally sub- sum；
The chitting piece number of germinating energy (%)=alloted days (3d)/for testing seed number × 100%；
Germination index=∑ (Gt/Dt), (in formula, the germinative number that Gt is t days, Dt is corresponding germination number of days)；
Relative germination rate (%)=(salt treatment germination percentage/control germination percentage) × 100%；
Opposite salt damage rate (%)=(control germination percentage-salt treatment germination percentage)/control germination percentage × 100%；
Accumulate germination percentage：G=N/S × 100%；N is the sum of chitting piece, and S is the seed number for experiment.
D, radiation effect of the radiation treatment to chitting piece under NaCl stress：
By radiation treatment seed different dose of radiations from carry out Seed Germination Test under different NaCl concentration gradients, 7 days
Statistics germination index, opposite salt damage rate and seedling percent afterwards, analysis various dose radiation group and different NaCl concentrations it
Between difference.
E, data processing and statistical method：
Data processing and graph making are carried out with Excel 2010, dual factors variance is carried out with 18.0 statistical softwares of SPSS
It analyzes and carries out significance of difference comparison with LSD methods.
In the step A and B, dose of radiation and different NaCl concentration gradients see the table below shown in 1.
Table 1
| Experimental group | Dose of radiation (Gy) | Time (min) | NaCl concentration gradient (mmolL-1) |
| 1 | 0 | 0 | 0、25、50、150、250、300 |
| 2 | 50 | 4 | 0、25、50、150、250、300 |
| 3 | 100 | 8 | 0、25、50、150、250、300 |
| 4 | 150 | 12 | 0、25、50、150、250、300 |
| 5 | 200 | 16 | 0、25、50、150、250、300 |
The diversiform-leaved poplar seed of the present invention60Co- gamma Rays method of mutagenesis utilizes60Co- γ carry out mutagenic treatment, in water planting
Mutagenic effect of the different dose of radiations with NaCl concentration gradient to diversiform-leaved poplar seed is observed under condition of culture.Determine best radiation agent
Amount.Establish diversiform-leaved poplar seed60The salt tolerant breeding technology of Co- gamma Rays mutagenesis.For the best dose of radiation of its radiaction mutation
Selection with NaCl concentration, which determines, provides foundation, and is the effective of current Desertification in Xinjiang, salinization of soil and secondary salinization soil
Using and exploitation, the natural regeneration to diversiform-leaved poplar forest vegetation artificial Populus Euphratica is built by seminal propagation, restore to be damaged ecological environment
Etc. have extremely important realistic meaning.It explores60Shadow of the Co- gamma Rays to Populus euphratica under Salt Stress Germination Characteristics
It rings, can significantly promote the sprouting of diversiform-leaved poplar seed under salt stress to seek60Co- gamma Rays dosage, to which radiation lures
Become breeding and provides new method and approach into diversiform-leaved poplar Germplasm enhancement and breed breeding.
It is research material that the present invention, which selects diversiform-leaved poplar seed, and radioinduction experiment is carried out to diversiform-leaved poplar seed.60Co- gamma-rays spokes
Penetrating breeding has many advantages, such as that mutation rate is high, the range of variation is big, therefore, uses60Co- gamma-rays carries out radiation to diversiform-leaved poplar seed and lures
Become, is conducive to high, salt tolerance and the higher diversiform-leaved poplar new varieties of breeding efficiency with germination percentage.
The diversiform-leaved poplar capsule of the lakes the Gan Jia of in September, 2015 Nature Reserve acquisition is as radioinduction material, by capsule
It is registered for numbering, capsule spontaneously dries, and seed villus sheds with its natural cracking, purer seed is obtained, under the conditions of 4 DEG C
The seed stored for future use is used as experiment material in 2016, and therefore, the germination percentage of seed is low, and seed very little, has to radiation
Higher sensibility.It selects diversiform-leaved poplar seed as radioactive breeding research object, is convenient for the culture dish culture item of different NaCl concentrations
Under part, seed has higher germination percentage, for by the way that successfully screening obtains salt tolerance seedling under the conditions of radioactive breeding bud rate occurred frequently
It lays the foundation.
Specific implementation process：
1, material：
It is national natural that diversiform-leaved poplar (Populus euphratica) seed is taken from the North SinKiang lakes preferred Natural Populus Euphratica Gan Jia
Protection zone.
2, method：
Condition of culture：
The processing of seed salt stress：Salinity is set as NaCl solution, concentration be disposed as 0 (CK), 25,50,150,250,
300mmol·L-1, totally 6 Salt Strees Condition processing, wherein 1 control treatment.
The above condition of culture is 25-27 DEG C of temperature, intensity of illumination is 3 000lx；Light dark period is 16/8h.
(2) radiation treatment
Diversiform-leaved poplar seed is test material, and grouping is used60Co- gamma-rays carries out radiation treatment；
60Co- gamma Rays dosage (0,50,100,150,200Gy), seed after radiation with 0.1% HgCl2It carries out
5~7min is sterilized, with 3 each 1min of distilled water flushing.Experiment sets 0 (CK, distilled water), 25,50,150,250 and altogether
300mmol·L-1NaCl concentration gradient, is taken 5mL processing solutions to pour into the culture dish of paving two layers of filter paper, and every 20 seeds connect
Kind is in culture dish, and totally 6 processing, every group of processing is repeated 4 times, is sealed and be placed in constant temperature illumination box with sealed membrane, temperature
It is cultivated under conditions of for 25-27 DEG C, intensity of illumination be 3 000lx, light dark cycles are 16/8h.Supplement single flash daily
Water is to ensure that each salinity is kept constant in Seed Germination.Every the sprouting situation for counting seed for 24 hours.(after inoculation
Start counting up within two days) statistics 7d.Determine the salt tolerant critical concentration that the seed of radioinduction germinates under salt stress.
(3) the germination index value of statistics seed, opposite salt damage rate and accumulation germination percentage：
Observation counts time and the chitting piece number of each dosage processing germination：Calculate germination index value with respect to salt damage rate and
Survival rate of seedling.
Germinate index determining：
Germination percentage (%)=(n/N) × 100%；N is the seed number that 7d normally germinates in formula, and N is for planting experimentally sub- sum；
The chitting piece number of germinating energy (%)=alloted days (3d)/for testing seed number × 100%；
Germination index=∑ (Gt/Dt), (in formula, the germinative number that Gt is t days, Dt is corresponding germination number of days)；
Relative germination rate (%)=(salt treatment germination percentage/control germination percentage) × 100%；
Opposite salt damage rate (%)=(control germination percentage-salt treatment germination percentage)/control germination percentage × 100%；
Accumulate germination percentage：G=N/S × 100%；N is the sum of chitting piece, and S is the seed number for experiment.
(4) radiation effect of the radiation treatment to chitting piece under NaCl stress
By radiation treatment seed different dose of radiations from carry out Seed Germination Test under different NaCl concentration gradients, 7 days
Statistics germination index, opposite salt damage rate and seedling percent afterwards, analysis various dose radiation group and different NaCl concentrations it
Between difference.
(5) data processing and statistical method
Data processing and graph making are carried out with Excel 2010, dual factors variance is carried out with 18.0 statistical softwares of SPSS
It analyzes and carries out significance of difference comparison with LSD methods.
Implement the final result of the present invention：
The irradiation of the various dose influence different to the germination generation of diversiform-leaved poplar seed from salinity processing.From fig. 1, it can be seen that hair
Bud rate, germinating energy and germination index (F=3.28, P between various dose radiation treatment<0.05；F=7.66, P<0.01；F=
19.965 P<0.01) (F=20.061, P between NaCl concentration<0.01；F=13.159, P<0.01；F=16.002, P<
0.01) otherness reaches the pole level of signifiance, shows germination of the seed under different dose of radiation processing in different salinity
There are larger differences for rate, germinating energy and germination index.Diversiform-leaved poplar seed exists60Co- gamma Rays to average germination percentage, germinating energy and
A variety of comparison results between germination index show (Fig. 3), in various dose radiation treatment, the germination under 150Gy processing
Rate (73.75 ± 1.25%, mean value ± SE), germination index (28.36 ± 1.21) and germinating energy (66.25 ± 2.39%) higher than pair
According to other processing groups, the significant difference (P compared with control group and 50Gy radiation treatments<0.05) it, but with 100 and 200Gy radiates
Processing (P not notable compared to difference>0.05).The average germination percentage of the lower seed of 50Gy processing is minimum (43.75 ± 8.5), and right
According to group and the difference of 200Gy dose of radiations without notable (P>0.05).
Fig. 2 is it is found that the radiation treatment of 100 and 150Gy is remarkably improved the germination percentage (P ＜ 0.05) of seed, with no spoke
It penetrates to handle to compare and 20% and 27.50% has been respectively increased；The radiation treatment of 200Gy improves 17.50%, but the not notable (P of difference
>0.05)；50Gy radiation treatments then inhibit the sprouting of seed, and germination percentage is made to reduce 2.5% (P>0.05) (Fig. 2).25,
50 and 150mmolL-1Under NaCl stress, the radiation of 50Gy significantly inhibits diversiform-leaved poplar germination, so that germination percentage is reduced respectively
23.75%, 36.25%, 41.25%, the processing of other dosage with compare between difference not significantly (Fig. 2).In 25mmolL- 1Under NaCl stress, germination percentage improves under the radiation treatment of 100 and 150Gy, and 15%, 16.25% has been respectively increased compared with control, poor
Different not significantly (P>0.05)；In 50mmolL-1Under NaCl stress, the facilitation of the radiation of 100 and 150Gy to germination
It is most apparent, 20%, 25% (P ＜ 0.05) has been respectively increased.150-300mmol·L-1Under NaCl stress, with NaCl concentration
Rise, the inhibiting effect that radiation treatment sprouts diversiform-leaved poplar seed is more notable, but 250mmolL-1Under NaCl stress, 50Gy dosage
Pole significantly improves 21.25% (p ＜ 0.01) (Fig. 2) to the germination percentage of radiation treatment compared with the control.
Percentage of seedgermination, germinating energy, opposite injury rate and relative germination rate under various dose radiation treatment is dense in NaCl
Otherness passes through pole significance test (P between degree<0.01).From the figure 3, it may be seen that under the processing of 150Gy doses, different NaCl
Germination percentage, germinating energy and relative germination rate under stress improve, and opposite salt damage rate reduces, according to the results of analysis of variance, in addition to 100Gy
Outside dose, average germination percentage, germinating energy, relative germination rate and opposite between salt damage rate and control group and other processing groups
Index value of germinateing difference extremely significantly (P ＜ 0.01), illustrates that the diversiform-leaved poplar seed sprouting under 150Gy doses coerce NaCl has one
Fixed facilitation.When dose of radiation is 50Gy, percentage of seedgermination, germinating energy and relative germination rate are substantially reduced (P ＜ 0.01),
Opposite salt damage rate increases (P ＜ 0.01).With the raising (100~200Gy) of dose of radiation, items germination index in it is raised become
Gesture, pole significant difference (P ＜ 0.01) compared with the control group, 200Gy and control group difference not significantly (P>0.05), with control group
(0Gy) is compared to 7.29%, 10.2% and 4.0% (Fig. 3) of increase.
Table 2 is shown as60Recurrence side of the Co- gamma Rays between diversiform-leaved poplar seed relative germination rate and different salinity
Journey.
Table 2
Wherein, * * indicate that, in 0.01 horizontal upper significant difference, wherein x indicates that salinity, Y indicate relative germination rate.
As shown in Table 2,60Exist between diversiform-leaved poplar seed relative germination rate and salinity under the processing of Co- gamma Rays notable
Negative correlativing relation (P<0.01).Regression equation between the relative germination rate handled by different dose of radiations and salinity is extrapolated
The salt tolerant suitable concentration (germination percentage reaches 75% NaCl concentration) that seed is sprouted, (germination percentage reaches 50% to salt tolerant critical concentration
NaCl concentration) and salt tolerant lethasl concentration (germination percentage be 10% NaCl concentration).
Under different dose of radiation processing, the salt tolerant suitable concentration of 100Gy and 150Gy seeds is 123~135mmolL-1,
Salt tolerant critical concentration is 172~181mmolL-1, its resistance to salinity high (dose of radiation CK groups, 50 compared with other dose of radiations
Salt tolerant suitable concentration and salt tolerant critical concentration with 200Gy are respectively 26~104mmolL-1, 119~159mmolL-1)。
An index of the salt tolerant critical concentration frequently as evaluation salt tolerance[18], according to the diversiform-leaved poplar seed under the processing of different dose of radiations
Salt tolerant critical concentration value (table 2), the salt tolerance (181mmolL of the seed under 150Gy processing-1) it is better than other radiation treatments
The salt tolerant critical concentration of seed, salt tolerance power are ordered as：150Gy>100Gy>200Gy>0Gy>50Gy.
In conclusion the sprouting favourable germination conditions of diversiform-leaved poplar seed are 150Gy doses and 181mmolL-1NaCl is dense
Degree.
The key problem in technology point of the present invention：
1, it using diversiform-leaved poplar seed as research material, uses60Co- gamma-rays carries out radiation treatment；
2, the radiation treatment to diversiform-leaved poplar seed is research shows that 181mmolL-1Under hypersaline environment, the processing of 150Gy doses
It can promote the sprouting of seed；
3, it is radiated under identified dosage, induced mutation rate is best, is conducive to the sprouting of diversiform-leaved poplar seed.
Finally it should be noted that：The foregoing is only a preferred embodiment of the present invention, is not intended to restrict the invention,
Although the present invention is described in detail referring to the foregoing embodiments, for those skilled in the art, still may be used
With technical scheme described in the above embodiments is modified or equivalent replacement of some of the technical features.
All within the spirits and principles of the present invention, any modification, equivalent replacement, improvement and so on should be included in the present invention's
Within protection domain.
Claims (3)
1. being used for the diversiform-leaved poplar seed of salt stress processing60Co- gamma Rays method of mutagenesis, which is characterized in that including following step
Suddenly：
1) condition of culture is determined：
The processing of diversiform-leaved poplar seed salt stress, concentration of sodium chloride solution 0-300mmolL are carried out using sodium chloride solution-1, diversiform-leaved poplar kind
The salt stress condition of culture of son is 23-27 DEG C of cultivation temperature, and intensity of illumination is 3 000lx, light dark period 16/8h；
2) radiation treatment：
By diversiform-leaved poplar seed with60Co- gamma-rays carries out radiation treatment,60Co- gamma Rays dosage is 0-200Gy, and radiated time is
0-16min；Seed after radiation with 0.1% HgCl2Carry out disinfection 5~7min, with distilled water flushing 3 times, each 1min, then
According to the concentration of sodium chloride solution of step 1), takes sodium chloride solution 5ml to pour into the culture dish of paving two layers of filter paper, 20 seeds are connect
Kind is repeated 4 times in culture dish, is sealed and be placed in constant temperature illumination box with sealed membrane, the culture item provided using step 1)
Part is cultivated；In incubation, first water is filled daily to ensure that each salinity is kept constant in Seed Germination,
Every the sprouting situation for counting seed for 24 hours, starts counting up within second day after inoculation, count 7d altogether.
2. the diversiform-leaved poplar seed according to claim 1 for salt stress processing60Co- gamma Rays method of mutagenesis,
It is characterized in that, in the salt stress processing in the step 1), concentration of sodium chloride solution 181mmolL-1, cultivation temperature 25-
27℃。
3. the diversiform-leaved poplar seed according to claim 2 for salt stress processing60Co- gamma Rays method of mutagenesis,
It is characterized in that, in the step 2),60Co- gamma Rays dosage is 150Gy, radiated time 0-16min.
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| CN110352850A (en) * | 2019-07-22 | 2019-10-22 | 广州中医药大学(广州中医药研究院) | A kind of method of the degeneration-resistant improved Varieties of breeding Herba Andrographitis high yield and high quality |
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| CN106332573A (en) * | 2016-08-26 | 2017-01-18 | 新疆林科院造林治沙研究所 | Method for promoting germination of populus euphratica seeds |
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