CN107047314A - A kind of method that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud - Google Patents
A kind of method that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud Download PDFInfo
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- CN107047314A CN107047314A CN201710405650.8A CN201710405650A CN107047314A CN 107047314 A CN107047314 A CN 107047314A CN 201710405650 A CN201710405650 A CN 201710405650A CN 107047314 A CN107047314 A CN 107047314A
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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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- 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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a kind of method for obtaining pure tetraploid winter jujube plant by blade callus induction and adventitious bud, this method sets up winter jujube tree excised leaf regenerating system, optimization minimal medium, plant growth regulator, AgNO3The conditions such as additives, set up chimera ' being stained with the winter No. 2 ' excised leaf high-efficiency regeneration system, inducer blade is grown thickly adventitious bud, the same method being combined using morphological index measure, chromosome number cytological observation and flow cytomery cell DNA, ploidy to leaf regeneration adventitious bud is identified, homozygosis tetraploid ' being stained with the winter No. 2 ' is filtered out, and it is carried out fast numerous.Tetraploid winter jujube plant is obtained by the purifying of natural bud mutation times sex mosaic, without undergoing transition culture, greenhouse production as in vitro tissue culture, transplanting the processes such as lower ground, crop field can be directly transplanted into, with the advantage of regrown material field Fast Evaluation can be carried out, the purification efficiency of ' being stained with the winter No. 2 ' is improved, accelerates to obtain pure tetraploid winter jujube.
Description
Technical field
The present invention relates to the mating system of plant, and in particular to a kind of to obtain pure four by blade callus induction and adventitious bud
The method of times body winter jujube plant.
Background technology
Jujube originates in China, is one of most characteristic and advantage fruit tree species of China, nearly 30,000,000 mu of cultivated area, yield
Many 6,000,000 tons, account for the world 99%.In recent years, with the development of jujube industry, the continuous adjusting and optimizing of cultivar structure, Zizyphus jujuba Mill
Cultivation is increasingly taken seriously.' winter jujube ' is the big main excellent Zizyphus jujuba Mill kind of cultivation of current China first, in addition to the Zhanhua of Shandong,
Also all reached a certain scale on Hebei, Shanxi and Shaanxi and other places.But ' winter jujube ' fruit is less than normal, cultivation management requires high, enters
Row ' winter jujube ' genetic improvement is not only ' winter jujube ', is also being necessarily required to for current Zizyphus jujuba Mill industry development.
Fruit polyploid often shows the merits such as the huge property of fruit, is constantly subjected to pay much attention in fruit breeding.
The polyploidy breeding of jujube tree starts from phase late 1970s, but jujube tree induced mutations polyploid is often acquisition chimera,
Chimera purifying is still the major issue for needing in jujube tree artificial polyploid mutagenesis to face.What current field large area was planted ' is stained with
Winter No. 2 ' it is derived from times sex mosaic material of ' winter jujube ' natural variation（2nd, four mixoplod）, its growth characteristics is unstable, ' the winter
Jujube ' fruit is less than normal, the annual diploid ' winter jujube ' that significant proportion occurs in branch, and its resistance is low, and freeze injury was tight in recent years
Weight, which increase management difficulty, reduces income, has had a strong impact on the popularization enthusiasm of jujube agriculture, limited the popularization of the kind
Development.In jujube tree ploidy germplasm innovation, existing induced mutations, endosperm culture obtain the report of polyploid, but natural bud mutation times
There is not been reported for the purifying research of sex mosaic.Can be by the purifying of natural bud mutation times sex mosaic, purifying chimera ' is stained with the winter
No. 2 ' tetraploid winter jujube is obtained, the polyploidization improvement and upgrading of regenerating of ' being stained with the winter No. 2 ' are realized, is urgent need to resolve in current industry
Technical problem.
The content of the invention
In order to solve above the problems of the prior art, the proposition of the invention it is a kind of by blade callus induction and
The method that adventitious bud obtains pure tetraploid winter jujube plant, this method obtains tetraploid by the purifying of natural bud mutation times sex mosaic
Winter jujube plant, without undergoing transition culture, greenhouse production as in vitro tissue culture, transplanting the processes such as lower ground, can directly be transplanted into
Crop field, with that can carry out the advantage of regrown material field Fast Evaluation, improves the purification efficiency of ' being stained with the winter No. 2 ', accelerates to obtain
Obtain pure tetraploid winter jujube.
The technical scheme that the application takes is as follows：
（1）The newborn axillary bud of ' being stained with the winter No. 2 ' jujube head branch or Secondary Branch terminal bud just sprouted in season of growth clip, are cut into 2-3
The cervical part of esophagus of individual blade, it is clean with aseptic water washing, then disinfected in alcohol on super-clean bench, it is finally clean with aseptic water washing,
Multiplying culture is carried out on MS culture mediums, every 5 weeks subcultures once, set up aseptic seedling cervical part of esophagus breeding system；
（2）From the blade of aseptic seedling top order clip Section 3 position 30d-40d leaf ages, entered in the way of vertical vane main lobe arteries and veins
Row cutting, it is impossible to penetrate between vein, otch at a distance of about 1-2mm, the blade handled well is contacted with adaxial and its surface the side of culture medium
Formula is inoculated on adventitious bud induction culture base, by the light culture of 2-4 weeks, then goes to optical culture；
（3）The Ploidy Identification of regenerated adventitious bud：Leaf index, internode carries out morphologic identification as index, utilizes routine
The chromosome number of pressed disc method identification induction plant, using fluorescence microscopy guard cell's number of chloroplast, gas rolls size
And density, the DNA content of plant cell is induced using flow cytomery, whether distinguish is that pure tetraploid winter jujube regeneration is indefinite
Bud.
（4）Pure tetraploid winter jujube regenerated adventitious bud, cultivates after 40d on adventitious bud induction culture base, blade is transferred to
Elongation culture is carried out on subculture medium, 20d is cultivated, more than 3cm adventitious bud is transferred on root media, root system is up to
After more than 4cm, de- bottle is transplanted into the pure tetraploid plantation in crop field.
（5）Potted plant resistant proof, the field resistance experiment of tetraploid plant are carried out, selection resistance level is high, character is excellent
Strain, the measure of fruit quality, so as to select excellent tetraploid winter jujube nursery stock.
Wherein above step（1）In MS culture mediums in added with 1.1 mg/L BA and 0.3mg/L IBA, culture temperature
Degree is 26+2℃。
Wherein above step（2）In adventitious bud culture base in be added with 1.5-2.0mg/L TDZ and 0.2-0.5mg/L
NAA and 5.0-7.0mg/L AgNO3。
Wherein above step（5）In the assay method of fruit quality be：The strain that antagonism level is high, character is excellent is entered
The measure of row fruit quality, the hardness of fruit is determined using GY-1 type disks hardometer, and the soluble solids content of fruit is utilized
ATAGO digital refractometers PRl is determined, and fruit Vitamin C content is determined using difference analysis spectrophotometer method, the size of fruit
Determined with weight, the presence or absence of fruit stone, so as to select excellent tetraploid winter jujube nursery stock.
' being stained with the winter No. 2 ' winter jujube tree described herein is two, tetraploid chimera.
Beneficial effects of the present invention are：The application obtains tetraploid winter jujube by the purifying of natural bud mutation times sex mosaic and planted
Strain, without undergoing transition culture, greenhouse production as in vitro tissue culture, transplanting the processes such as lower ground, can directly be transplanted into crop field, have
There is the advantage that can carry out regrown material field Fast Evaluation, improve the purification efficiency of ' being stained with the winter No. 2 ', accelerate to obtain pure four times
Body winter jujube.
The application is to be stained with No. 2 times of sex mosaics of winter as material, by combining preferred field, blade callus induction and adventitious bud
Obtain method purifying two, ' being stained with the winter No. 2 ' of four times of mixoplods, the excellent homozygosis winter jujube of acquired character of pure tetraploid winter jujube plant
Tetraploid plant simultaneously promotes demonstration, promotes the replacement upgrading of winter jujube kind, improves the yield and quality of winter jujube, individual tree volume increase
At least 50%.Technical scheme is from application value of material importance and novelty, technical advance and result of study etc.
From the point of view of aspect, compared with the correlative study in the domestic and international industry, advanced level is in.
Embodiment
The following examples are that the present invention is described in further detail.
Embodiment：
1. project implementation place
The field seed selection of winter jujube nursery stock is carried out in the Chinese new and high technology seedling-wood breeding of Zhanhua County winter jujube research institute subordinate.
2. project implementation scale
The different phase of project research, scale is ascending, and the scale of every kind of material is at 1000 square metres or so, to the development phase
Popularization.
3. project implementation method
（1）Season of growth clip just sprouted two, the newborn axillary bud of tetraploid chimera ' being stained with the winter No. 2 ' jujube head branch or Secondary Branch top
Bud, is cut into the cervical part of esophagus with 2-3 blade, clean with aseptic water washing, is then disinfected in alcohol on super-clean bench, finally with nothing
Bacterium water is rinsed well, and Multiplying culture is carried out on the BA and 0.3mg/L IBA MS culture mediums added with 1.1 mg/L, culture
Temperature is 26+2 DEG C, every 5 weeks subcultures once, set up aseptic seedling cervical part of esophagus breeding system；
（2）From the blade of aseptic seedling top order clip Section 3 position 30d-40d leaf ages, entered in the way of vertical vane main lobe arteries and veins
Row cutting, it is impossible to penetrate between vein, otch at a distance of about 1-2mm, the blade handled well is contacted with adaxial and its surface the side of culture medium
Formula is inoculated in the NAA and 5.0-7.0mg/L of TDZ and 0.2-0.5mg/L added with 1.5-2.0mg/L AgNO3Adventitious bud
On inducing culture, by the light culture of 2-4 weeks, optical culture is then gone to；
（3）The Ploidy Identification of regenerated adventitious bud：Leaf index, internode carries out morphologic identification as index, utilizes routine
The chromosome number of pressed disc method identification induction plant, using fluorescence microscopy guard cell's number of chloroplast, gas rolls size
And density, the DNA content of plant cell is induced using flow cytomery, whether distinguish is that pure tetraploid winter jujube regeneration is indefinite
Bud.
Leaf index is about 1.85, averagely about 3.7 centimetres of internode, and the chromosome for inducing plant is identified using conventional compression
Number, chromosome number is 48, and tetraploid is measured as using flow cytometer.
（4）Pure tetraploid winter jujube regenerated adventitious bud, cultivates after 40d on adventitious bud induction culture base, blade is transferred to
Elongation culture is carried out on subculture medium, 20d is cultivated, more than 3cm adventitious bud is transferred on root media, root system is up to
After more than 4cm, de- bottle is transplanted into the pure tetraploid plantation in crop field.
（5）Potted plant resistant proof, the field resistance experiment of tetraploid plant are carried out, selection resistance level is high, character is excellent
Strain, the measure of fruit quality, sample 100kg, and measurement result is as follows：
The hardness of fruit is determined using GY-1 type disks hardometer, and average hardness is up to 8.5 ㎏ ∕ c ㎡, the soluble solids content of fruit
Determined using ATAGO digital refractometers PRl, soluble solid is 32%, fruit Vitamin C content uses difference analysis light splitting
Photometer measuring method is determined, and Vitamin C content is 314 ㎎ ∕ 100g, and the size and weight of fruit are determined, mean fruit weight 21.52g, most
Big single fruit weight 57.6g, fruit shape index is 0.912, and fruit stone is small, so as to select excellent tetraploid winter jujube nursery stock.
Claims (5)
1. a kind of method that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud, it is characterised in that step is such as
Under：
（1）The newborn axillary bud of ' being stained with the winter No. 2 ' jujube head branch or Secondary Branch terminal bud just sprouted in season of growth clip, are cut into 2-3
The cervical part of esophagus of individual blade, it is clean with aseptic water washing, then disinfected in alcohol on super-clean bench, it is finally clean with aseptic water washing,
Multiplying culture is carried out on MS culture mediums, every 5 weeks subcultures once, set up aseptic seedling cervical part of esophagus breeding system；
（2）From the blade of aseptic seedling top order clip Section 3 position 30d-40d leaf ages, entered in the way of vertical vane main lobe arteries and veins
Row cutting, it is impossible to penetrate between vein, otch at a distance of about 1-2mm, the blade handled well is contacted with adaxial and its surface the side of culture medium
Formula is inoculated on adventitious bud induction culture base, by the light culture of 2-4 weeks, then goes to optical culture；
（3）The Ploidy Identification of regenerated adventitious bud：Leaf index, internode carries out morphologic identification as index, utilizes routine
The chromosome number of pressed disc method identification induction plant, using fluorescence microscopy guard cell's number of chloroplast, gas rolls size
And density, the DNA content of plant cell is induced using flow cytomery, whether distinguish is that pure tetraploid winter jujube regeneration is indefinite
Bud；
（4）Pure tetraploid winter jujube regenerated adventitious bud, cultivates after 40d on adventitious bud induction culture base, blade is transferred into subculture
Elongation culture is carried out on culture medium, 20d is cultivated, more than 3cm adventitious bud is transferred on root media, root system is up to 4cm
After above, de- bottle is transplanted into the pure tetraploid plantation in crop field；
（5）Potted plant resistant proof, the field resistance experiment of tetraploid plant are carried out, selection resistance level is high, the strain that character is excellent
System, the measure of fruit quality, so as to select excellent tetraploid winter jujube nursery stock.
2. a kind of side that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud as claimed in claim 1
Method, it is characterised in that：Step（1）In MS culture mediums in added with 1.1 mg/L BA and 0.3mg/L IBA, cultivation temperature
26+2℃。
3. a kind of side that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud as claimed in claim 1
Method, it is characterised in that：Step（2）In adventitious bud culture base in TDZ and 0.2-0.5mg/L added with 1.5-2.0mg/L
NAA and 5.0-7.0mg/L AgNO3。
4. a kind of side that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud as claimed in claim 1
Method, it is characterised in that the assay method of fruit quality is：The strain that antagonism level is high, character is excellent carries out the survey of fruit quality
Fixed, the hardness of fruit is determined using GY-1 type disks hardometer, and the soluble solids content of fruit utilizes ATAGO digital refractometers
PRl is determined, and fruit Vitamin C content is determined using difference analysis spectrophotometer method, and the size and weight of fruit are determined, fruit stone
The presence or absence of, so as to select excellent tetraploid winter jujube nursery stock.
5. a kind of side that pure tetraploid winter jujube plant is obtained by blade callus induction and adventitious bud as claimed in claim 1
Method, it is characterised in that：Described ' being stained with the winter No. 2 ' winter jujube tree is two, tetraploid chimera.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115011634A (en) * | 2022-07-07 | 2022-09-06 | 安徽农业大学 | Genetic transformation method for promoting wild jujube callus to form adventitious buds by using phytoplasma effector SJP 1and SJP2 |
| CN115530076A (en) * | 2022-11-10 | 2022-12-30 | 山东滨州国家农业科技园区管理服务中心(滨州黄河三角洲高效生态产业现代技术研究院) | Winter jujube tissue culture method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105918117A (en) * | 2016-05-30 | 2016-09-07 | 沾化县冬枣研究所 | Propagation method of tetraploid winter jujube |
| CN105941154A (en) * | 2016-05-30 | 2016-09-21 | 沾化县冬枣研究所 | Comprehensive breeding method for superior winter jujube seedlings |
| CN106034608A (en) * | 2016-05-30 | 2016-10-26 | 沾化县冬枣研究所 | Cultivating method for improving yield of jujube tree |
| CN106688881A (en) * | 2016-12-16 | 2017-05-24 | 北京林业大学 | Method for separating homozygotes from ploidy chimera plant |
-
2017
- 2017-06-01 CN CN201710405650.8A patent/CN107047314A/en not_active Withdrawn
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105918117A (en) * | 2016-05-30 | 2016-09-07 | 沾化县冬枣研究所 | Propagation method of tetraploid winter jujube |
| CN105941154A (en) * | 2016-05-30 | 2016-09-21 | 沾化县冬枣研究所 | Comprehensive breeding method for superior winter jujube seedlings |
| CN106034608A (en) * | 2016-05-30 | 2016-10-26 | 沾化县冬枣研究所 | Cultivating method for improving yield of jujube tree |
| CN106688881A (en) * | 2016-12-16 | 2017-05-24 | 北京林业大学 | Method for separating homozygotes from ploidy chimera plant |
Non-Patent Citations (2)
| Title |
|---|
| 王娜等: "秋水仙素离体诱导冬枣和酸枣四倍体", 《园艺学报》 * |
| 黄建等: "枣树离体叶片不定芽再生体系建立的研究", 《西北植物学报》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115011634A (en) * | 2022-07-07 | 2022-09-06 | 安徽农业大学 | Genetic transformation method for promoting wild jujube callus to form adventitious buds by using phytoplasma effector SJP 1and SJP2 |
| CN115011634B (en) * | 2022-07-07 | 2024-01-30 | 安徽农业大学 | Genetic transformation method for promoting wild jujube callus to form adventitious buds by using phytoplasma effector SJP 1and SJP2 |
| CN115530076A (en) * | 2022-11-10 | 2022-12-30 | 山东滨州国家农业科技园区管理服务中心(滨州黄河三角洲高效生态产业现代技术研究院) | Winter jujube tissue culture method |
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Application publication date: 20170818 |