CN102293152A - Combined germination method of hull peeling off and low temperature seed soaking for Bothriochloa ischaemum (L.) Keng - Google Patents
Combined germination method of hull peeling off and low temperature seed soaking for Bothriochloa ischaemum (L.) Keng Download PDFInfo
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- CN102293152A CN102293152A CN 201110191912 CN201110191912A CN102293152A CN 102293152 A CN102293152 A CN 102293152A CN 201110191912 CN201110191912 CN 201110191912 CN 201110191912 A CN201110191912 A CN 201110191912A CN 102293152 A CN102293152 A CN 102293152A
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Abstract
The invention relates to a combined germination method of hull peeling off and low temperature seed soaking for Bothriochloa ischaemum (L.) Keng. The method comprises two parts: the method of hull peeling off is employed to completely remove hulls of seeds of Bothriochloa ischaemum (L.) Keng so as to destroy hard physical structure of hulls and break dormancy of seeds of Bothriochloa ischaemum (L.) Keng; the method of low temperature seed soaking is employed for rapid improvement of germination capacity of Bothriochloa ischaemum (L.) Keng. The method specifically comprises four steps: (1) careful choosing and removing of dopants; (2) low temperature seed soaking; (3) removing of dopants in seeds; (4) germination of seeds. According to the invention, germination capacity of seeds without hulls in the invention is increased from 45% to more than 90% compared to seeds with hulls for contrast; accordingly, the invention is of important practical significance and extensive applicable value to cultivation of and experimental research on seeds of Bothriochloa ischaemum (L.) Keng.
Description
Technical field
The invention belongs to biological technical field, relate to the germinating method of plant seed breaking dormancy, specifically is exactly to break the yellow bluestem Seed Dormancy fast, significantly improves the combined method of white poplar grass seed germination rate on the MS solid culture medium.
Background technology
Yellow bluestem [Bothriochloa ischaemum (L.) Keng] is a grass family Bothriochloa herbaceos perennial, is a kind of good forage.This herbage originates from south of europe and Asia, and all there is distribution in each continent now.Mainly be distributed in North China, south, northwest, Loess Plateau and low mountains and hills district, the middle and south, Shanxi in China, for hilly and mountainous land is mainly herded grass seeds.Yellow bluestem has the very high economic worth and the ecological value.Yellow bluestem begins to be food that various domestic animal is liked from sprouting, and You Yiyang likes food the most.Analyze according to China Agricultural University, the yellow bluestem protein content belong to medium more than, digestibility is higher.Protein can match in excellence or beauty with the pulse family grass before the heading.Yellow bluestem grows thickly, the tool stolon, has fixing of soil and conservation of water, the drought resisting anti-erosion, and vitality is strong, advantages such as anti-trample.Yet in recent years, because environmental deterioration and overgraze cause original yellow bluestem meadow to suffer great destruction.Therefore, how to enlarge the task of top priority that the yellow bluestem cultivated area has become production and scientific research fast.
The yellow bluestem seed has the biological property of dormancy.Wild yellow bluestem seed exists germination rate and the low shortcoming of vigor
[1], wild seed generally only has 10～20% low germination rate.This laboratory test finds that also yellow bluestem is induced when germinateing on the MS solid culture medium, the percentage of seedgermination ratio is broadcast when soil also low, and in other herbages, as also having found this type of phenomenon in the sheep's hay.Studies show that in the past, the yellow bluestem seed has to a certain degree requirement to low temperature, need be similar to the freezing in advance of vernalization; Shi Weiwei
[1]Deng the influence of having inquired into PEG-6000 dialogue germination of guineagrass seed, experimental result finds that germination rate, germination vigor, germination index and the vitality index of the PEG-6000 solution initiation dialogue Leymus chinensis seeds of the various flows of water all have no significant effect.But the flow of water causes the speed of germination that can effectively improve seed for the PEG-6000 solution of-0.6Mpa.Tang Long
[2]Inquired into the influence of dialogue germination of guineagrass seed such as light, pH, temperature, found that the sprouting of light dialogue Leymus chinensis seeds does not have influence, but natural lighting promptly day alternates with night can promote every index of its sprouting; The sprouting of alkaline environment (7.0≤pH≤8.0) the dialogue Leymus chinensis seeds of appropriateness has facilitation; ≤ 7 ℃ of dialogue germination of guineagrass seed produce inhibitory action, also occur inhibition to a certain degree more than 32 ℃, and 27 ℃ are the suitableeest seed germination temperature.
Summary of the invention
The present invention by arranging the control of technology and temperature to combine in advance, creates a kind of combination germinating method in order to improve the yellow bluestem percentage of seedgermination, and it comprises following a few part: (1) seed cleaning impurity elimination: the yellow bluestem seed grain is little, and maturity is inconsistent, and ripening rate is low.After the wild yellow bluestem seed results, need selected impurity elimination.(2) go bran to handle: the yellow bluestem seed has the bran skin hard, that quality is fine and close, and water penetration, gas permeability are relatively poor, and the mechanical-physical obstacle of bran skin is having a strong impact on the suction and the germination of yellow bluestem seed.Therefore, be Ying's shell with the bran skin of yellow bluestem seed, all peel off removal, purpose is to destroy the firm physical arrangement of bran skin, removes the yellow bluestem seed dormancy to a certain extent, can significantly improve the yellow bluestem germination rate.With the seed bran method of going in the technology of arranging in advance the bran skin portion of Leymus chinensis seeds is all removed, purpose is to destroy the firm physical arrangement of bran skin.(3) germinating method: temperature is the important factor that influences seed sprouting.Generally speaking, suitable temperature environment promotes seed germination, the too high or too low seed germination that all suppresses.Utilize the low-temperature seed presoaking technology to improve the yellow bluestem percentage of seedgermination fast.The ingenious combination of above-mentioned several parts has breaks the yellow bluestem seed dormancy fast, significantly improves the effect of yellow bluestem percentage of seedgermination.
The present invention specifically realizes through the following steps:
1. selected impurity elimination: select the seed of brown, full, regular shape, reject khaki, slight of stature, seed in irregular shape
[2]
2. low-temperature seed presoaking: with the seed of selected impurity elimination, remove the bran skin after, immerse fully in the sterile water, place under 0 ℃～5 ℃, dark condition soaked 24 hours;
Wherein, the temperature of preferred low-temperature seed presoaking is 0 ℃.
3. seed disinfection: the seed that will soak through low temperature, soak 30s with absolute alcohol earlier, use sterile water wash again 3～4 times, soak 15min with the 0.1w% mercuric chloride solution afterwards, use sterile water wash again 7～8 times;
4. seed sprouting: the seed after sterilization, be linked in the MS solid culture medium, 40 seeds of each culture dish inoculation place under 25～30 ℃, and dark condition was cultivated 15～20 days.
Beneficial effect: the present invention can make the yellow bluestem germination rate be brought up to more than 90% by 40% by the bran normal temperature processing seed germination rate that promptly has of contrast.Therefore, dialogue Leymus chinensis seeds of the present invention experimental study and cultivation all have important practice significance and value widely in particular for tissue culture and transgenic research.
Embodiment
Following non-limiting example can make those of ordinary skill in the art more fully understand the present invention, but does not limit the present invention in any way.
The yellow bluestem seed that this experiment is used derives from high bridge township, Ansai County, Yan'an, the Shaanxi Province area bamboo yellow bluestem meadow, adret slope of collapsing.
Absolute alcohol and mercuric chloride are all available from Sangon Biotech (Shanghai) Co., Ltd..
MS (Murashige and Skoog, 1962) solid culture based component is (mg/L of unit):
Macroelement: NH
4NO
3: 1650; KNO
3: 1900; CaCl
22H
2O:440; MgSO47H
2O:370; KH
2PO
4: 170; Trace element: KI:0.83; H
3BO
3: 6.2; MnSO
44H
2O:22.3; ZnSO
47H
2O:8.6; Na
2MnO
42H
2O:0.25; CuSO
45H
2O:0.025; CoCl
26H
2O:0.025; Molysite: FeSO
47H
2O:27.8; Na
2-EDTA2H
2O:37.3; Organic substance: inositol: 100; Nicotinic acid: 0.5; Vitamin B6: 0.5; Vitamin B1: 0.1; Glycine: 2.0, agar: 8g; Solvent is a deionized water.
Embodiment 1
(1) selected impurity elimination
The ratio of mature seed and ateliosis seed is 3: 1 in the wild yellow bluestem seed of gathering
[2]Therefore at first adopt artificial method that seed is carried out selected impurity elimination.Select the mature seed of brown, full, regular shape, reject the seed of khaki, slight of stature, ateliosis in irregular shape.From 1100 wild yellow bluestem seeds, choose 800.
(2) go bran to handle
Method is all peeled the bran skin (being Ying's shell) of 800 yellow bluestem seeds off by hand, in order to carrying out the low-temperature seed presoaking of seed.
(3) low-temperature seed presoaking
A. will go the yellow bluestem seed of bran to put into the test tube of 15ml respectively, totally 3 test tubes, each test tube is put into 40 seeds, in each test tube, pour the sterile water of 10ml into, with the complete submergence of seed, 3 test tubes are put into 0 ℃, 5 ℃, 28 ℃ low temperature refrigerator respectively, under the dark condition, soaked 24 hours.
Repeating step a three times.
B. will put into the test tube of 15ml without the yellow bluestem seed that the past bran is handled respectively, totally 3 test tubes, each test tube is put into 40 seeds, in each test tube, pour the sterile water of 10ml into, with the complete submergence of seed, 3 test tubes are put into 0 ℃, 5 ℃, 28 ℃ low temperature refrigerator respectively, under the dark condition, soaked 24 hours.
Repeating step b three times.
(4) seed disinfection
For the seed soaking seeds treated in above-mentioned 18 test tubes, respectively water is outwelled earlier, absolute alcohol with 10ml soaks 30s again, pours out alcohol, uses the sterile water wash seed of 10ml again, repeat to use sterile water wash 3 times, 0.1w% mercuric chloride solution with 10ml soaks 15min afterwards, pours out the 0.1w% mercuric chloride solution, with the sterile water wash seed of 10ml, repeat to carry out seed germination experiment with after the sterile water wash 8 times.
(5) germination test
Seed after the above-mentioned sterilization is linked into respectively in the MS solid culture medium, and 40 seeds of each culture dish inoculation are inoculated 18 culture dishes altogether, under the dark condition, carry out constant temperature culture 20 days at 28 ℃, calculate germination rate, germination vigor, statistics, as table 1:
Table 1 low-temperature seed presoaking is handled mainly the germinate influence of index of dialogue Leymus chinensis seeds with removing bran
Wherein:
Germination rate=chitting piece number/always test seed number;
Germination vigor=germination begins chitting piece number in the peak period of germinateing/always test seed number;
Germination vigor calculates with the germination rate that germinateed the 7th day, all the other testing results for germinateing the 20th day.
The experimental result that obtains is as shown in table 1.The yellow bluestem seed that removes bran under three kinds of Temperature Treatment all than under the same terms bran percentage of seedgermination height being arranged.This may be owing to go bran to destroy the firm physical arrangement of bran skin, has promptly broken Seed Dormancy, helps seed again simultaneously and absorbs moisture.Experiment finds that also along with the raising of soaking the seed temperature, no matter be the seed of bran and the seed that does not remove bran, germination rate reduces along with the rising of temperature, shows the similar vernalization of yellow bluestem seed germination needs, and low temperature soaks and helps the yellow bluestem seed germination.Overall target on germination rate, germination vigor, the average germinating time, with seeds palea removing, 0 ℃ soak 24 hours, in the MS solid culture medium, carry out the dark germination combination resultant effect the best of cultivating of 28 ℃ of constant temperature.
The germination vigor that goes seven day the germination rate of yellow bluestem seed under 0 ℃ of condition of bran to reach 98.2%, the 20 day is 21.3%, compares with the data 44.7% that control group does not go the seed of bran to obtain at normal temperatures, has nearly improved 50%.Under uniform temp, go the yellow bluestem seed germination rate of bran also all to be greatly improved than the seed that does not remove bran.Show that bran and low-temperature seed presoaking broken the dormancy of yellow bluestem fast, improved the germination rate of yellow bluestem fast.
Wild yellow bluestem seed exists germination rate and the low shortcoming of vigor
[1]The data of this experiment contrast group show too, even under the condition of moisture and illumination abundance, the seed germination rate at normal temperatures that does not remove bran only is 45% low germination rate, will not go the seed of bran to germinate at normal temperatures after depositing 10 hours under-25 degree again, and its germination rate only is about 80%
[2]The present invention can make the yellow bluestem germination rate be brought up to more than 98% by 45% by the bran normal temperature processing seed germination rate that promptly has of contrast.Therefore, dialogue Leymus chinensis seeds of the present invention experimental study and cultivation all have important practice significance and value widely in particular for tissue culture and transgenic research.
List of references:
[1] the wide tiger of Dong Shi Weiwei etc., PEG-6000 causes the influence of yellow bluestem seed sprouting, grassland and lawn [J], 2007, (6): 26-28.
[2] Tang Long, four kinds of grass seeds Water Physiology ecological characteristic researchs of one's native land of northern Shensi Loess Region [D], Shaanxi: northwest agricultural university, 2005.
Claims (2)
1. a yellow bluestem removes bran and low-temperature seed presoaking combination germinating method, it is characterized in that step is:
A. selected impurity elimination: select the seed of brown, full, regular shape, reject khaki, slight of stature, seed in irregular shape;
B. low-temperature seed presoaking: the seed that will handle through above-mentioned steps a, remove the bran skin after, immerse fully in the sterile water, place under 0 ℃～5 ℃, dark condition soaked 24 hours;
C. seed disinfection: through above-mentioned steps b seeds treated, soak 30s with absolute alcohol earlier, use sterile water wash again 3～4 times, soak 15min with the 0.1w% mercuric chloride solution afterwards, use sterile water wash again 7～8 times;
D. seed sprouting: through above-mentioned steps c seeds treated, be linked in the MS solid culture medium, 40 seeds of each culture dish inoculation place under 25～30 ℃, and dark condition was cultivated 15～20 days.
2. yellow bluestem according to claim 1 removes bran and low-temperature seed presoaking combination germinating method, it is characterized in that, the temperature of low-temperature seed presoaking is 0 ℃ among the described step a.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668766A (en) * | 2012-05-10 | 2012-09-19 | 黑龙江省科学院高技术研究院 | Germination method of Leymus chinensis seeds |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101199251A (en) * | 2006-12-14 | 2008-06-18 | 中国科学院东北地理与农业生态研究所 | Leymus chinensis seeds palea removing and temperature changing combination germinating method |
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2011
- 2011-07-08 CN CN 201110191912 patent/CN102293152A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101199251A (en) * | 2006-12-14 | 2008-06-18 | 中国科学院东北地理与农业生态研究所 | Leymus chinensis seeds palea removing and temperature changing combination germinating method |
Non-Patent Citations (3)
| Title |
|---|
| 《中国优秀博硕士学位论文全文数据库（硕士） 农业科技辑》 20060215 唐龙 "陕北黄土丘陵区四种乡土草种水分生理生态特征研究" 第57-59、63、68页 1-2 , 第2期 * |
| 《草业科学》 20031231 朱宇旌等 "牧草种子休眠解除方法综述" 第24-27页 1-2 第20卷, 第3期 * |
| 《草原与草坪》 20071231 史威威等 "PEG-6000引发对白羊草种子发芽的影响" 第26-28页 1-2 , 第6期 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102668766A (en) * | 2012-05-10 | 2012-09-19 | 黑龙江省科学院高技术研究院 | Germination method of Leymus chinensis seeds |
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Application publication date: 20111228 |