CN109197573A - A kind of rapid breeding method of high yield and high quality oil tea - Google Patents
A kind of rapid breeding method of high yield and high quality oil tea Download PDFInfo
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- CN109197573A CN109197573A CN201811271779.5A CN201811271779A CN109197573A CN 109197573 A CN109197573 A CN 109197573A CN 201811271779 A CN201811271779 A CN 201811271779A CN 109197573 A CN109197573 A CN 109197573A
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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
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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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- 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 present invention provides a kind of rapid breeding methods of high yield and high quality oil tea, method includes the following steps: the 1) screening of breeding parent；2) breeding parent is hybridized, harvests F1 generation cenospecies；3) F1 generation cenospecies anther Fiber differentiation；4) callus differentiation culture；5) it is good for seedling rooting culture；6) rooting culture 7) Field planting.The present invention combines anther culture technique with conventional cross-breeding means, breeding practice applied to oil tea, oil tea training regeneration plant is obtained for the first time, and the oil tea new lines of high yield and high quality have successfully been obtained by further oil tea economic indicator and the screening of the consaturated oil index of quality.Compared with the breeding cycle of simple crossbreeding many decades, method of the invention greatly shortens the breeding time limit, breeding efficiency is improved, has greatly accelerated oil tea breeding process, is all of great significance to the breeding of oil tea breeding and popularization and camellia oleiferaindustry healthy and rapid development.
Description
Technical field
The present invention relates to nursery stock breeding fields, and in particular to a kind of rapid breeding method of high yield and high quality oil tea.
Background technique
Oil tea in the narrow sense refer to C. olelfera (Camellia oleiferaAbel.) also known as ribes tree, tea oil tree,
White flower tea, be Theaceae (Theaceae) Camellia (Camellia) one kind, main morphological features be dungarunga or shrub, flower
White, basidixed, capsule, because its seed can extract oil (tea oil) it is edible due to gain the name.And sensu lato oil tea is Theaceae Camellia
Seed oil content is higher in plant, and the general designation for the tree species for having certain cultivation to manage area is also wrapped not only including C. olelfera
Include camellia meiocarpa, Camellia yuhsienensis, C.semiserrata, Camellia Vietnamensis, Gaozhou camellia, narrow leaf oil tea, Camellia Vietnamensis, bolt shell Camellia,
The types such as Camellia chekiangoleosa, Nanrong oil tea, Tengchong safflower oil tea, Camellia nitidissima.Oil tea likes sunshine, is afraid of cold, it is desirable that year
16 ~ 18 DEG C of temperature on average, it is mainly distributed on China Yangtze river basin and south area, in addition, in Burma, Vietnam, Malaysia and day
Also there is a small amount of distribution in this grade state.China is that the maximum oil tea place of production, existing camellia oleifera lam area are about 4,000,000 hm in the world2,
Wherein the provinces such as Hunan, Jiangxi, Guangxi, Fujian, Zhejiang, Guangdong are the oil tea main producing regions in China.
Oil tea and oil palm, olive and coconut and the referred to as big traditional oil tree in the world four have high comprehensive utilization
Value.The most important effect of oil tea is oil expression first, therefore the oil content of oil tea, fatty acid composition are to measure tea-oil tree yield and matter
The important indicator of amount.It is known as tea oil using the oil that Seed of Camellia oleifera is squeezed out, tea oil is a kind of good edible oil, it is full of nutrition,
Rich in unsaturated fatty acid and the essential trace elements of the human body, it is known as " east olive oil ".The unsaturated fatty acid of tea oil contains
Amount is up to 90% or more, based on oleic acid and linoleic acid, also containing the high prices unsaturated fatty acid such as a small amount of linolenic acid, and without pair
The harmful erucic acid of human body, it is deep to be liked by the masses.Tea oil has preferable health care and medical function, and wherein it is total can to reduce blood for oleic acid
Cholesterol and bad cholesterol, do not reduce good cholesterol but, and oleic acid is called " safe fats acid " by nutrition circle；Linoleic acid is people
Body cannot synthesize but be required unsaturated fatty acid, it is the primary raw material of synthesis of prostaglandins, and have reduction gallbladder solid
Pure and mild blood lipid improves the specific functions such as hemorheological property and its prevention coronary heart disease；Tea oil mid-oleic is relatively stable, linoleic acid
Content amplitude of fluctuation is larger, thus oleic acid content number can be used as evaluation tea oil quality one of the important signs that.Secondly oil tea
It is all chemical industry, light industry, food, feed work containing Tea Saponin, tea seed polysaccharide, tea seed albumen etc. in the leached tea oil slag left after oil expression
The raw material of industry product etc.；Third oil tea is a kind of evergreen, long-lived tree species, primary to plant, and harvest time is general to plant more than a century
8-10 closing afterwards are grown into forest, and not only can increase oil sources, but also afforestation rate can be improved, and are had and are beautified the environment, conserve water and soil, conserving water
Source, the ecological functions regulated the climate.
Although the comprehensive utilization value of oil tea is very high, the camellia oleiferaindustry in China obtains enough development not yet, master
It wants the reason is that the universal yield of oil tea is not high, and yield is unstable, is unable to satisfy the bigger market demand.According to statistics, China's tea oil
About 200,000 tons of average annual output, and wherein 70% yield from 30% plant, it is seen that low yield weak plant proportion is quite big.Oil tea
, there are various aspects in the reason of low yield, and as Seedling propagation traditionally causes, standing forest variet complexity, weak plant ratio are big, and strain produces difference
Greatly；Many camellia oleifera lam stand structure are unreasonable, dilute close unevenness；Mostly for same hall, standing forest aging, natural renovation ability difference etc., still
Most basic reason is that oil tea genetic variation and genetic differentiation is serious, and germplasm is mixed and disorderly, very different, and breeding level is low.Therefore it is produced to increase oil tea
Amount improves oil tea quality, accelerates the breeding paces of high-yield tea-oil fine quality, it is current oil tea that the whole improved variety of raising is horizontal
Top priority in production.
All the time, conventional cross-breeding is responsible for key player in camellia oleifera cultivar breeding.Crossbreeding is that one kind changes
Good plant Parent for Quality Traits creates the important and effective approach of new variety of plant, and the hybrid generation generated by hybridization is in addition to can
With the merit of hereditary parents, the organism of hybrid generation can also be caused to generate hereditary variation, to show some new
Merit, these new merits can often overcome or weaken some original bad characters of its hybrid parent.
But some problems existing for conventional cross-breeding also constrain the development of oil tea breeding technique, such as oil to a certain extent
Tea is xylophyta, and breeding cycle is long, and the time of many decades will be spent by carrying out breeding of new varieties；Oil tea belongs to cross-pollination plant
Object is constantly in height heterozygous state, is difficult to be sheerly by selfing means, and breeding progeny characteristic properties complexity without
Stablize, individual plants are very different.Therefore there is an urgent need to be reformed for the breeding technique of oil tea.
Haploid breeding is one of modern biotechnology breeding technique, can use in breeding F1 generation cenospecies anther,
Pollen or unfertilized ovary obtain monoploid material by vitro culture and double to generate double single times through nature or artificial chromosome
Body, dihaploid (DH) have the characteristic of completely homozygous homogeneity and are the self-mating systems of standard, therefore not by haploid breeding
The separation of hybrid character can be only overcome to shorten the breeding time limit, and can be with rapid breeding self-mating system, to greatly speed up breeding
Process.Haploid breeding can also select excellent work to offer convenience for breeding, and the double haploid obtained after doubling monoploids can
It excludes to interfere caused by the recessive factor of remarkable indicator cover in heterozygote, the phenotype of plant is consistent with genotype, improves choosing
The accuracy and reliability selected.If haploid breeding technology combined with conventional breeding means, the new product applied to oil tea
In kind breeding, there will be great development prospect.
Vitro anther culture is the material base of the haploid most common means of artificial induction and haploid breeding.So
And the domestic research about oiltea anther culture at present is few, and most researchs only rest on Anther Culture and induce callus group
It knits, only grand vibration hero once had C. olelfera to obtain callus by Anther Culture and formed the report of green bud point and root, this says
Bright oiltea anther culture still has problems in practice process.There are many influence factor that Anther Culture is subject to, main to wrap
Include genotype, the stage of development of anther, pretreatment, the influence of cultural method and the condition of culture, " bottle of oiltea anther culture at present
Neck problem " is that callus how to be made to differentiate complete regeneration plant.Years of researches pass through in team of the present invention, preliminary to establish
Oiltea anther culture regenerating system, and oil tea training regeneration plant has successfully been obtained, finally realize high-yield tea-oil high-quality product
The rapid breeding of system.
Summary of the invention
The purpose of the present invention is making improvement and innovation for shortcoming and defect present in background technique, provide a kind of high
The rapid breeding method of high-quality oil tea is produced, this method substantially reduces the breeding time limit of oil tea breeding, has high practical valence
Value.
In order to achieve the above-mentioned object of the invention, technical solution provided by the invention is as follows:
A kind of rapid breeding method of high yield and high quality oil tea, which is characterized in that method includes the following steps:
1) it the screening of breeding parent: according to the breeding objective of high yield and high quality, filters out consistent excellent with breeding objective with more
Two camellia oleifera cultivars that benign shape and suitable locality are planted/it is as breeding parent；
2) breeding parent is hybridized, harvests F1 generation cenospecies: at the florescence of breeding parent, the side pollinated using conventional manual
Method hybridizes the two, harvests F1 generation cenospecies after next year is solid；By the positive season plantation of F1 generation cenospecies in crop field；
3) it F1 generation cenospecies anther Fiber differentiation: when F1 generation cenospecies growth and development enters flowering stage, acquires and filters out small spore
Son development enters the healthy bud of mid-late uninucleate stage, bud is first placed in 4 DEG C of 5 ~ 7d of Cold pretreatment, then at cold plasma
15 ~ 20s is managed, bud carries out surface sterilization by treated, then carefully strips anther (excision filigree) and is inoculated into Fiber differentiation
On base, 40 ~ 50d of culture induces callus under conditions of 23 ~ 25 DEG C of temperature, dark；
4) callus differentiation culture: when above-mentioned callus size it is long to 2 ~ 3mm when, be forwarded on differential medium in time,
55 ~ 65d is cultivated under conditions of 24 ~ 26 DEG C of temperature, 1400 ~ 1800lx of intensity of illumination, 10 ~ 12h/d of light application time, it is during which every
28d or so subculture 1 time, callus can differentiate seedling；
5) it is good for seedling rooting culture: when the Seedling Height that above-mentioned steps differentiate grows to 3 ~ 4cm, being transferred to strong seedling rooting culture solution
In, 26 ~ 32d, which is cultivated, under conditions of 24 ~ 26 DEG C of temperature, 2000 ~ 2400Lx of intensity of illumination, light application time 13-15h/d is given birth to
Long healthy and strong flower training regeneration plant；
6) rooting culture: after flower training regeneration plant grows 3 ~ 5 true leaves, 6 ~ 8 root systems, being transferred to temperature is 15 ~ 30 DEG C, humidity
To tame 7 ~ 10d under the conditions of natural lighting in 80 ~ 90% greenhouse, flower training regeneration plant is taken out from culture bottle then, is moved
It plants into the cultivation matrix in greenhouse, conventional cultivation management；
7) Field planting: when the flower training regeneration plant plant height of above-mentioned greenhouse production grows to 20cm or so, as oil tea kind
Seedling is colonized to big Tanaka, is normally applied fertilizer, deinsectization, diseases prevention and tending management, to the development of oil tea seedling early growth into the manhood
Afterwards according to the oil tea strain of oil tea economic indicator and high-quality index of quality further screening high yield and high quality.
High yield and high quality specifically refers to have single plant yield height, dry seed-producing rate according to oil tea economic indicator in the step 1)
The characteristic that high, fresh seed-producing rate is high, kernel oil content is high has unsaturated fatty acid total content high, oily according to oil quality index
The high characteristic of acid content.
The method that screening microspore development period is in the bud of mid-late uninucleate stage in the step 3) are as follows: will be in bud
2 ~ 3 pieces of anther picking are pulverized, and pollen is released, and are dyed by DAPI and are determined that Pollen stage is mid-late uninucleate stage.
Cold plasma is handled in the step 3) method particularly includes: in vacuum sealing environment, using helium as work
Medium, the Non-ionizing radiation for carrying out 15 ~ 20s to oil tea bud under the processing power of 60W are handled.
The method of bud surface sterilization in the step 3) are as follows: bud is rinsed well with tap water, and is blotted with filter paper
Moisture, then aseptically, first with 75% alcohol disinfecting 60s, aseptic water washing 2 times, then with 0.1% mercuric chloride sterilize
15min, aseptic water washing 5 times.
The ingredient of induced medium in the step 3) are as follows: MS culture medium+3.1 ~ 3.7mg/L+ of tryptophan mannitol 14 ~
2.6 ~ 3.2mg/L+4-IPA of 20g/L+ soybean lecithin 4.0 ~ 4.6mg/L+ lanthanum nitrate 2.3 ~ 2.7mg/L+ glycine betaine 0.9 ~
1.1mg/L+2,4-D 0.4 ~ 0.6mg/L+ polyvinylpyrrolidone 15 ~ 22g/L+ oil tea shell powder 3.5 ~ 4.5g/L, pH be 5.4 ~
5.8。
The ingredient of differential medium in the step 4) are as follows: KNO31400 ~ 1800mg/L, NH4NO3850 ~ 900mg/L,
KH2PO4330 ~ 360mg/L, MgSO4·7H2O 144 ~ 162mg/L, CaCl2·2H2O 375 ~ 400mg/L, MnSO4·4H2O
20 ~ 25mg/L, zinc methionine 8.5 ~ 9.5mg/L, H3BO37.8 ~ 8.6mg/L, KI 0.6 ~ 0.8mg/L, CuSO4·5H2O
0.023 ~ 0.027mg/L, CoCl2·6H2O 0.023 ~ 0.027mg/L, Na2MoO4·2H2O 0.20 ~ 0.30mg/L, FeSO4·
7H2O 27 ~ 28mg/L, Na236 ~ 37mg/L of-EDTA, 85 ~ 95mg/L of inositol, vitamin B6 0.5 ~ 0.6mg/L, NaN31.4~
1.8 mg/L, 0.5 ~ 0.6mg/L of vitamin B1,0.3 ~ 0.4mg/L of beta carotene, D-VB5 calcium 3.1 ~ 3.7mg/L, 5'- flesh
Thuja acid 7.5 ~ 8.0mg/L of disodium, 50 ~ 60mg/L of mashed peas, 2.2 ~ 2.8mg/L of polyglutamic acid, tetramethyl glutaric acid 0.4 ~
0.6mg/L, N- phenyl 0.8 ~ 1.2mg/L of phthalamidic acid, 1.1 ~ 1.3mg/L of compound sodium nitrophenolate, Pollen Astragali Melilotoidis (Pollen Astragali Sinici) 20 ~
28mg/L, 3.6 ~ 4.4g/L of camellia seed oil, 28 ~ 34g/L of maltose, plant gel 5 ~ 7g/L, pH are 5.4 ~ 5.8.
Be good in the step 5) seedling rooting culture solution ingredient be 1/2MS culture medium (be free of agar)+silicon Feng Huan 0.8 ~
1.0mg/L+NAA0.1 ~ 0.3mg/L+ 36 ~ 40mg/L of mashed peas, pH value is 5.6 ~ 6.0.
Beneficial effects of the present invention:
1) present invention establishes complete oiltea anther culture body by carrying out exploratory development to oiltea anther culture process for the first time
System, and oil tea training regeneration plant has successfully been obtained.
2) present invention combines anther culture technique with conventional cross-breeding means, applied to the breeding practice of oil tea,
And the oil tea new lines of high yield and high quality have successfully been filtered out, the oil tea breeding time limit is greatly shortened, oil tea breeding effect is improved
Rate is all of great significance to the breeding of oil tea breeding and popularization and camellia oleiferaindustry healthy and rapid development.
Specific embodiment
It is carried out below in conjunction with rapid breeding method of the specific embodiment to high yield and high quality oil tea provided by the invention detailed
Explanation, but they cannot be interpreted as limiting the scope of the present invention.
Embodiment 1
A kind of rapid breeding method of high yield and high quality oil tea, method includes the following steps:
1) screening of breeding parent: according to high yield and high quality, (single plant yield is high, dry seed-producing rate is high, fresh seed-producing rate is high, kernel oil content
It is high, oleic acid content is high) breeding objective, it is suitable for the long woods of C. olelfera kind No. 3 and Zhejiang in Zhejiang plantation that we, which filter out,
Green field 25 is used as breeding parent.No. 3 fruit sizes of long woods are medium, and sexennial plant single fruit yield about 3.0kg does seed
Rate is 24.0%, and kernel oil content is 46.8%, and yield is more stable.The kernel oil content 52.1% of Qingtian County of Zhejiang 25, unsaturated fat
Acid content 90.6%, oleic acid content 81.3%, oil quality is excellent.
2) breeding parent is hybridized, harvest F1 generation cenospecies: 2007, in the flower of long woods No. 3 and Qingtian County of Zhejiang 25
Phase is hybridized the two using the method that conventional manual pollinates, and harvests F1 generation cenospecies after next year is solid；2008 by F1 generation
The positive season plantation of cenospecies is in crop field.
3) F1 generation cenospecies anther Fiber differentiation: when F1 generation cenospecies growth and development enters flowering stage within 2012, acquisition is simultaneously
Filter out the healthy bud (method of screening are as follows: by 2 ~ 3 pieces of anther picking in bud that microspore development enters mid-late uninucleate stage
Pulverize, release pollen, dyed by DAPI and determine that Pollen stage is mid-late uninucleate stage), bud is first placed in 4 DEG C of low temperature
6d is pre-processed, then is handled with cold plasma, concrete operation method are as follows: in vacuum sealing environment, using helium as working media,
The Non-ionizing radiation for carrying out 18s to bud under the processing power of 60W is handled, and by treated, bud is rinsed well with tap water,
And with filter paper suck dry moisture, then aseptically, first with 75% alcohol disinfecting 60s, aseptic water washing 2 times, then with 0.1%
Mercuric chloride sterilize 15min, aseptic water washing 5 times, then carefully strip anther (excision filigree) and be inoculated into induced medium,
40 ~ 50d of culture induces callus under conditions of 24 DEG C of temperature, dark, and counts callus induction rate；
The ingredient of induced medium are as follows: MS culture medium+tryptophan 3.4mg/L+ mannitol 17g/L+ soybean lecithin 4.3mg/L+
Lanthanum nitrate 2.5mg/L+ glycine betaine 2.9mg/L+4-IPA 1.0mg/L+2,4-D 0.5mg/L+ polyvinylpyrrolidone 18.5g/L
+ oil tea shell powder 4.0g/L, pH 5.6.
4) callus differentiation culture: when above-mentioned callus size it is long to 2 ~ 3mm when, be forwarded to differential medium in time
On, under conditions of 25 DEG C of temperature, intensity of illumination 1600lx, light application time 11h/d cultivate 55 ~ 65d, during which every 28d or so after
In generation 1 time, callus can differentiate the seedling of green, and count plantlet differentiation rate；
The ingredient of differential medium are as follows: KNO31600mg/L, NH4NO3875mg/L, KH2PO4345mg/L, MgSO4·7H2O
153mg/L, CaCl2·2H2O 387.5mg/L, MnSO4·4H2O 22.5mg/L, zinc methionine 9.0mg/L, H3BO3 8.2mg/
L, KI 0.7mg/L, CuSO4·5H2O 0.025mg/L, CoCl2·6H2O 0.025mg/L, Na2MoO4·2H2O 0.25mg/
L, FeSO4·7H2O 27.5mg/L, Na2- EDTA 36.5mg/L, inositol 90mg/L, vitamin B6 0.55mg/L,
NaN31.6mg/L, vitamin B1 0.55mg/L, beta carotene 0.35mg/L, D-VB5 calcium 3.4mg/L, 5'-inosinic acid two
Sodium 7.7mg/L, mashed peas 55mg/L, polyglutamic acid 2.6mg/L, tetramethyl glutaric acid 0.5mg/L, N- phenyl neighbour's carboxybenzoyl
Amine 1.0mg/L, compound sodium nitrophenolate 1.2mg/L, Pollen Astragali Melilotoidis (Pollen Astragali Sinici) 24mg/L, camellia seed oil 4.0g/L, maltose 31g/L, plant are solidifying
Glue 6g/L, pH 5.6.
5) it is good for seedling rooting culture: when the Seedling Height that above-mentioned steps differentiate grows to 3 ~ 4cm, being transferred to strong seedling rooting training
In nutrient solution, 26 ~ 32d is cultivated under conditions of 25 DEG C of temperature, intensity of illumination 2200Lx, light application time 14h/d, obtains robust growth
Flower training regeneration plant；
The ingredient of strong seedling rooting culture solution is 1/2MS culture medium (being free of agar)+silicon Feng Huan 0.9mg/L+NAA0.2mg/L+ pea
Beans mud 38mg/L, pH value 5.8.
6) rooting culture: flower training regeneration plant grow 3 ~ 5 true leaves, 6 ~ 8 root systems after, be transferred to temperature be 15 ~ 30 DEG C,
In the greenhouse that humidity is 80 ~ 90%, 7 ~ 10d is tamed under the conditions of natural lighting, then takes flower training regeneration plant from culture bottle
Out, it is transplanted in the cultivation matrix in greenhouse, conventional cultivation management.
7) Field planting: when the flower training regeneration plant plant height of above-mentioned greenhouse production grows to 20cm or so, as oil
Tea planting seedlings are normally applied fertilizer, deinsectization, diseases prevention and tending management, 2015 start, to oil tea seedling early growth to big Tanaka
Develop the oil tea after entering the manhood according to oil tea economic indicator and unsaturated fatty acid index further screening high yield and high quality
Strain.
Embodiment 2
The influence for inducing and breaking up to prove cold plasma preprocess method of the invention to oiltea anther, the present embodiment take
The cold plasma to bud in 1 step 3) of the embodiment that disappeared pre-processes, and bud is only placed in 4 DEG C of Cold pretreatment 6d,
He operates same as Example 1, statistics callus induction rate and plantlet differentiation rate.
Embodiment 3
In order to prove influence that differential medium of the invention breaks up oiltea anther, the present embodiment will be in 1 step 4) of embodiment
The differential medium of use replaces with N6+2.0mg/LBA+0.5mg/LIAA+0.5mg/L5Br+1.0mg/L riboflavin, the differentiation
Culture medium is male " oiltea anther culture obtains green bud point and root " from grand vibration, counts plantlet differentiation rate.
Interpretation of result
1, the influence that different pretreatments method induces oiltea anther and breaks up, embodiment 1 and the callus of embodiment 2 are lured
Conductance and plantlet differentiation rate statistics such as following table one.
The cold plasma preprocess method that the present invention uses it can be seen from upper table result can be improved oiltea anther training
Callus induction rate and plantlet differentiation rate in supporting, it may be possible to oil tea bud be pre-processed using cold plasma technology, energy
Pollen viability and germination are enough improved, histocyte Differentiation is promoted.
2, the plantlet differentiation rate of embodiment 1 and embodiment 3 is united in the influence that different differential mediums break up oiltea anther
Meter such as following table two.
Differential medium of the invention is used it can be seen from upper table result, plantlet differentiation rate has reached 8.5%, hence it is evident that high
In embodiment 3.It mentions, is not obtained due to pollution complete green in documents " oiltea anther culture obtains green bud point and root "
Seedling, and 1.2% plantlet differentiation rate is obtained in the test of embodiment 3, it may be possible to due to not polluted in test, and
The influence generated before using preprocess method of the invention and anther Fiber differentiation.Differential medium Reasonable Regulation And Control of the invention
The proportion of basic element is more suitable for oiltea anther differentiation culture, also added NaN3, beta carotene, D-VB5 calcium, pea
Mud, polyglutamic acid, tetramethyl glutaric acid, N- phenyl phthalamidic acid, 5'-inosinic acid disodium, compound sodium nitrophenolate, Chinese milk vetch flower
The substances such as powder, camellia seed oil can enhance cell changes, promote callus differentiation.
Oil tea is obtained in embodiment 1 and trains 73 plants of regeneration plant, according to oil tea economic indicator and unsaturated fatty acid index
The further screening oil tea strain of 2 high yield and high quality.
The technical principle of the invention is described above in combination with a specific embodiment.These descriptions are intended merely to explain of the invention
Principle, and shall not be construed in any way as a limitation of the scope of protection of the invention.Based on the explanation herein, the technology of this field
Personnel can associate with other specific embodiments of the invention without creative labor, these modes are fallen within
Within protection scope of the present invention.
Claims (8)
1. a kind of rapid breeding method of high yield and high quality oil tea, which is characterized in that method includes the following steps:
1) it the screening of breeding parent: according to the breeding objective of high yield and high quality, filters out consistent excellent with breeding objective with more
Two camellia oleifera cultivars that benign shape and suitable locality are planted/it is as breeding parent；
2) breeding parent is hybridized, harvests F1 generation cenospecies: at the florescence of breeding parent, the side pollinated using conventional manual
Method hybridizes the two, harvests F1 generation cenospecies after next year is solid；By the positive season plantation of F1 generation cenospecies in crop field；
3) it F1 generation cenospecies anther Fiber differentiation: when F1 generation cenospecies growth and development enters flowering stage, acquires and filters out small spore
Son development enters the healthy bud of mid-late uninucleate stage, bud is first placed in 4 DEG C of 5 ~ 7d of Cold pretreatment, then at cold plasma
15 ~ 20s is managed, bud carries out surface sterilization by treated, then carefully strips anther (excision filigree) and is inoculated into Fiber differentiation
On base, 40 ~ 50d of culture induces callus under conditions of 23 ~ 25 DEG C of temperature, dark；
4) callus differentiation culture: when above-mentioned callus size it is long to 2 ~ 3mm when, be forwarded on differential medium in time,
55 ~ 65d is cultivated under conditions of 24 ~ 26 DEG C of temperature, 1400 ~ 1800lx of intensity of illumination, 10 ~ 12h/d of light application time, it is during which every
28d or so subculture 1 time, callus can differentiate seedling；
5) it is good for seedling rooting culture: when the Seedling Height that above-mentioned steps differentiate grows to 3 ~ 4cm, being transferred to strong seedling rooting culture solution
In, 26 ~ 32d, which is cultivated, under conditions of 24 ~ 26 DEG C of temperature, 2000 ~ 2400Lx of intensity of illumination, light application time 13-15h/d is given birth to
Long healthy and strong flower training regeneration plant；
6) rooting culture: after flower training regeneration plant grows 3 ~ 5 true leaves, 6 ~ 8 root systems, being transferred to temperature is 15 ~ 30 DEG C, humidity
To tame 7 ~ 10d under the conditions of natural lighting in 80 ~ 90% greenhouse, flower training regeneration plant is taken out from culture bottle then, is moved
It plants into the cultivation matrix in greenhouse, conventional cultivation management；
7) Field planting: when the flower training regeneration plant plant height of above-mentioned greenhouse production grows to 20cm or so, as oil tea kind
Seedling is colonized to big Tanaka, is normally applied fertilizer, deinsectization, diseases prevention and tending management, to the development of oil tea seedling early growth into the manhood
Afterwards according to the oil tea strain of oil tea economic indicator and high-quality index of quality further screening high yield and high quality.
2. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 1)
Middle high yield and high quality specifically refers to have single plant yield height, dry seed-producing rate height, fresh seed-producing rate height, kernel according to oil tea economic indicator
The high characteristic of oil content, according to oil quality index, the characteristic high with unsaturated fatty acid total content, oleic acid content is high.
3. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 3)
The method that middle screening microspore development period is in the bud of mid-late uninucleate stage are as follows: 2 ~ 3 pieces of anther picking in bud are pulverized,
Pollen is released, is dyed by DAPI and determines that Pollen stage is mid-late uninucleate stage.
4. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 3)
Middle cold plasma processing method particularly includes: in vacuum sealing environment, using helium as working media, the processing power of 60W
Under to oil tea bud carry out 15 ~ 20s Non-ionizing radiation handle.
5. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 3)
The method of middle bud surface sterilization are as follows: rinse bud with tap water well, and with filter paper suck dry moisture, then in aseptic condition
Under, first with 75% alcohol disinfecting 60s, aseptic water washing 2 times, then with 0.1% mercuric chloride sterilize 15min, aseptic water washing 5 times.
6. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 3)
The ingredient of middle induced medium are as follows: MS culture medium+tryptophan 3.1 ~ 3.7mg/L+, 14 ~ 20g/L+ of mannitol soybean lecithin 4.0 ~
0.9 ~ 1.1mg/L+2,4-D of 4.6mg/L+ lanthanum nitrate 2.3 ~ 2.7mg/L+, 2.6 ~ 3.2mg/L+4-IPA of glycine betaine 0.4 ~
0.6mg/L+ polyvinylpyrrolidone 15 ~ 22g/L+ oil tea shell powder 3.5 ~ 4.5g/L, pH are 5.4 ~ 5.8.
7. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 4)
The ingredient of middle differential medium are as follows: KNO31400 ~ 1800mg/L, NH4NO3850 ~ 900mg/L, KH2PO4330 ~ 360mg/L,
MgSO4·7H2O 144 ~ 162mg/L, CaCl2·2H2O 375 ~ 400mg/L, MnSO4·4H220 ~ 25mg/L of O, zinc methionine
8.5 ~ 9.5mg/L, H3BO37.8 ~ 8.6mg/L, KI 0.6 ~ 0.8mg/L, CuSO4·5H20.023 ~ 0.027mg/L of O,
CoCl2·6H2O 0.023 ~ 0.027mg/L, Na2MoO4·2H2O 0.20 ~ 0.30mg/L, FeSO4·7H227 ~ 28mg/L of O,
Na236 ~ 37mg/L of-EDTA, 85 ~ 95mg/L of inositol, vitamin B6 0.5 ~ 0.6mg/L, NaN31.4 ~ 1.8 mg/L, vitamin
0.5 ~ 0.6mg/L of B1,0.3 ~ 0.4mg/L of beta carotene, D-VB5 3.1 ~ 3.7mg/L of calcium, 5'-inosinic acid disodium 7.5 ~
8.0mg/L, 50 ~ 60mg/L of mashed peas, 2.2 ~ 2.8mg/L of polyglutamic acid, tetramethyl glutaric acid 0.4 ~ 0.6mg/L, N- phenyl are adjacent
Carboxybenzoyl 0.8 ~ 1.2mg/L of amine, 1.1 ~ 1.3mg/L of compound sodium nitrophenolate, 20 ~ 28mg/L of Pollen Astragali Melilotoidis (Pollen Astragali Sinici), camellia seed oil 3.6 ~
4.4g/L, 28 ~ 34g/L of maltose, plant gel 5 ~ 7g/L, pH are 5.4 ~ 5.8.
8. a kind of rapid breeding method of high yield and high quality oil tea according to claim 1, which is characterized in that the step 5)
In be good for seedling rooting culture solution ingredient be 1/2MS culture medium (be free of agar)+0.8 ~ 1.0mg/L+NAA0.1 of silicon Feng Huan ~
0.3mg/L+ 36 ~ 40mg/L of mashed peas, pH value are 5.6 ~ 6.0.
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| CN111903277A (en) * | 2020-08-24 | 2020-11-10 | 山东省种子有限公司 | Seed treatment method for improving carrot root expansion speed and application thereof |
| CN118255984A (en) * | 2024-04-08 | 2024-06-28 | 临沂中磷生物科技有限公司 | Preparation method, device and application of rooting-promoting polyglutamic acid derivative |
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