CN104109032B - A kind of culture medium increasing cotton fiber length and cultural method thereof - Google Patents
A kind of culture medium increasing cotton fiber length and cultural method thereof Download PDFInfo
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Abstract
The present invention relates to cotton fiber field, particularly to a kind of culture medium increasing cotton fiber length and cultural method thereof.A kind of culture medium increasing cotton fiber length, adds second component, any one in following material of second component: brassinosteroid, (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, brassinosteroid and (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, sucrose, potassium chloride, mannitol in BT culture medium.A kind of culture medium increasing cotton fiber length that the present invention provides, use cotton ovule in-vitro culture method, eliminate the complexity that Herb participates in, eliminate under field condition, the change of moisture, illumination, temperature etc., control condition that can be artificial in vitro preferably studies the development mechanism of cotton fiber；The length of the cotton fiber obtained by this culture medium culturing is greatly improved, and this is significant to the raising of cotton quality.
Description
Technical field
The present invention relates to cotton fiber field, increase cotton fiber in particular to one
The culture medium of length and cultural method thereof.
Background technology
Cotton fiber is to be extended by the epidermis cell of ovule external integument to form, long fibre and chopped fiber
Be all one unicellular, ripe cotton fiber is flat tubular, by base portion, middle part, top
Composition.Cotton fiber is made up of primary cell wall, secondary cell wall and lumen from outside to inside, nascent born of the same parents
Wall is the prototype structure of fiber, is made up of pectin, and there is waxiness outside；Secondary cell wall is by fiber
Element is constituted, in wheel stricture of vagina shape level.
Cotton fiber development terminates from blooming to blow-of-cottons, typically at 40-50 days, according to its physiology generation
Thank to difference, cotton fiber development process is divided into the differentiation of fibre initial cells, elongation, secondary
Wall thickeies and ripe four the continuous developmental stages of dehydration.The differentiation of cotton fiber germinal cell refers to
Ovule epidermis cell is differentiated to form fibre initial cells, and 3 days before Cotton Gossypii blooms start, open
Spent completed the same day.Fibre initial cells intracytoplasmic IAA oxidase is broken by acid subsequently
Badly cause IAA activity to be improved, promote epidermis cell projection, and then promote fiber
Elongation, within this period, the fiber of early differentiation forms long fibre, late differentiation
Fiber forms chopped fiber, typically spends latter 30 days fibers to reach greatest length.The elongate fiber phase and
There is the overlap period of a very long time the secondary wall thickening phase, and cotton fiber dehydration maturation is from splitting bell
To abundant blow-of-cottons, about about 5 days, after cotton boll splits, fiber dewatering was dried, cell death,
Shorten flat tube shape into, owing to fibre bundle deposits in the shape of a spiral, the shadow of the internal force because being produced by dehydration
Ring so that filament contraction turns bent, final fiber maturation.
Cotton fiber development in addition to being affected by genotype, the environment such as temperature, moisture and illumination
Factor and cultivation condition all have a strong impact on Fibre Development, and then affect cotton fibre quality.Base
Because type is self-evident on the impact of cotton fiber quality.In environmental factors, temperature is restriction
Cotton Fiber Differentiation and the principal element of growth, temperature is high, and elongation is fast, and the time is short, otherwise,
Temperature is low, then elongating stage is long.Moisture is indirect by affecting the growth of cotton plant and physiological process
Affecting cotton fiber development, illumination is then mainly to affect secondary wall and the deposit of cellulose and affect
Fiber quality.
The method utilizing high-flux sequence, to lint-free fuzzless lintless mutant and the transcript profile of wild type
It is analyzed, discovery signal transduction pathway, lipid metabolic pathways, auxin signal pathway,
Invertase signal approach, the approach such as Fiber Metabolism approach, phosphatase and dehydrogenase or gene are at fibre
Dimension grows its important function in early days.
White jade woodss etc., with new land early No. 16 materials such as grade, determine IAA during cotton fiber development
With the content of the endogenous hormones such as GA, result show the fiber rapid elongation phase IAA and
The content of the hormones such as GA is higher, thus it is speculated that the elongation of cotton fiber is had necessarily by IAA and GA etc.
Facilitation.The differentiation of ovule epidermis cell and IAA content have substantial connection, Post flowering
0-3d, IAA content is the highest, and ovule epidermal differentiation is the most, and directly increases cotton seed surface
Raw fiber count.Exogenous hormone IAA and GA all can effectively facilitate the elongation of fiber, but
The two effect stage is different, and wherein IAA mainly acts on the elongating stage of cotton fiber, and GA
Then mainly act on the early stage of cotton fiber development.A Zhao Jing ancient type of banner hoisted on a featherdecked mast and Wang Longhua (2000) research find,
Under the conditions of suitable pH, the elongation of cotton fiber is all had by GA3, GA4 and GA4+7
Significantly facilitation, but every kind of gibberellins has a suitable concentration, and each not phase
With.IAA, GA and KT are in vitro fiber initial and the impact of growth, and result shows individually
Use KT can not promote the growth of fiber, and can promote when IAA with GA is used alone
The growth of fiber, and when using, facilitation becomes apparent from simultaneously.Tan Xia ridge (2009) measures
In Post flowering 6d cotton fiber and ovule, the content of vegetable oil black alcohol, finds that it is fast at fiber
Speed has higher content elongating stage, declines at secondary wall synthesis phase then content.At In vitro Embryo
The BT culture medium that pearl is cultivated adds the black alcohol of vegetable oil, and fibre length the most substantially shortens,
Thus speculate that the black alcohol of vegetable oil and elongate fiber have substantial connection.External source second is dilute also can be promoted in vitro
The growth of fiber.
Plant tissue culture is one of current most widely used biotechnology.Ovule development is
Study the important method that in vitro fiber growth is grown.Ovule development refers to separate from ovary
Ovule, and it is inoculated in culture medium the process cultivated.Cotton ovule isolated culture is that Cotton Gossypii is fine
The research of dimension growth promoter provides experiment porch, and some that can avoid the Nature are uncertain
The impact of factor (weather, temperature, illumination etc.), and realize a large amount of steady production cotton fiber and stretch
One of long important method grown, can be as research cell elongation and a reference of differentiation
Model.
Between cotton quality supervision and inspection center of the Ministry of Agriculture was to China 1998-2011 14 years
The main breed in main product cotton region has carried out the investigation and analysis of fiber quality characteristics and has pointed out that China is raw
It is interval that product field cotton fiber upper half mean length is mainly distributed on center line 28.0-30.9mm,
Account for 77.1%；Fiber length uniformity index reaches at or above top grade (83.0%-85.9%)
Sample accounts for 55.57%.As can be seen here, the fiber length uniformity in Cotton in China fiber quality
Preferably, and fibre length only based on medium staple, it is difficult to fully meet middle short flannel and the need of middle long wool
Asking, improve Cotton in China fiber quality, particularly adjusting cotton fiber length distribution becomes me
The key issue that state's cotton breeding is urgently to be resolved hurrily.
Summary of the invention
It is an object of the invention to provide a kind of culture medium increasing cotton fiber length and training thereof
Breeding method, to solve above-mentioned problem.
Provide a kind of culture medium increasing cotton fiber length in an embodiment of the present invention,
Second component is added, described second component appointing in following material in BT culture medium
A kind of: brassinosteroid, (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, brassinosteroid and (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, sucrose,
Potassium chloride, mannitol.
Preferably, described second component is brassinosteroid, the content of described brassinosteroid
For 0.05-1 μm ol/L.
Preferably, the content of described brassinosteroid is 0.1-0.5 μm ol/L.
Preferably, described second component is (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate
For 0.05-7 μm ol/L.
Preferably, the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5-5 μm ol/L.
Preferably, described second component is brassinosteroid and (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, described Brassica campestris L
The content of element lactone is 0.05-0.2 μm ol/L, and the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5-5
μmol/L。
Preferably, described second component is sucrose, and the content of described sucrose is 2-15g/L.
Preferably, described second component is potassium chloride, and the content of described potassium chloride is 0.01-0.08
mol/L。
Preferably, described second component is mannitol, and the content of described mannitol is 3-10
g/L。
Preferably, described BT culture medium is possibly together with heteroauxing and gibberellins；Described indole
The addition of acetic acid is 8-12 μm ol/L, and the addition of described gibberellins is 4-6 μm ol/L.
Present invention also offers a kind of cultural method increasing cotton fiber length, including following
Step: field Cotton Gossypii blooms and carries out selfing listing mark the previous day, takes Post flowering three days
The ovule of children's bell, then uses the cotton fiber length that increases described in any one of right 1-9
Culture medium carries out in vitro light culture；Cultivation temperature is 30 DEG C ± 2 DEG C.
A kind of culture medium increasing cotton fiber length that the embodiment of the present invention provides, adopts
Use cotton ovule in-vitro culture method, eliminate the complexity that Herb participates in, eliminate
Under field condition, the change of moisture, illumination, temperature etc., permissible in vitro
Artificial control condition preferably studies the development mechanism of cotton fiber；By this culture medium
The length cultivating the cotton fiber obtained is greatly improved, and this is to cotton quality
Improve significant.
Detailed description of the invention
Below by specific embodiment, the present invention is described in further detail.
Embodiment 1
1.1 test sites are originally tested in January, 2014-May cotton in the Chinese Academy of Agricultural Sciences
Hainan scientific research center of flower institute Cotton Gossypii National Key Laboratory's biology is carried out.
1.2 test material materials to be tested are palm fibre 1-61, palm fibre 263, RT white cotton fiber, green cotton CC28,
Green cotton G88, all material Jun You the Chinese Academy of Agriculture Science and Technologys Cotton Research Institute's national middle term storehouse carries
Supply.All material has the annual selfing of this laboratory to preserve.The plant growth regulator of experiment
In having heteroauxing (IAA), gibberellins (GA3), (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate (MeJA) and Brassica campestris L element
Ester (BR) is purchased from Aladdin.Potassium chloride, mannitol and sucrose are domestic analytical pure.Embryo
The chemical reagent of pearl isolated culture is public purchased from Amresco company of Germany or U.S. Sigma
Department.
The configuration of BT culture medium: BT culture medium prescription is as shown in table 1.By the thing in formula
After plasmogamy makes, pH is adjusted to 5.0, and BT culture medium used is divided in 100ml conical flask
In, every bottle adds 50ml culture medium, through autoclaving (121 DEG C, 20min), standby.
Table 1BT culture medium prescription
1.3 test method
1.3.1 Ovule development
Field Cotton Gossypii bloom the previous day time carry out selfing listing mark；In 8:00--9 in the morning:
00, take Post flowering and carry out ovules culture in vitro the young age of three days, bud first peelled off by the young bell of ovule
After leaf and sepal again with 70% ethanol to ovary sterilize 3～4min；After sterilization with 95% wine
The essence infiltration 2-3 second；Then it is put into sterilized culture dish after alcohol exposure lamp flame envelope, treats fire
Flame carefully separates ovule with tip tweezers after extinguishing；Ovule is inoculated in the cultivation of different disposal
On base, it is positioned over light culture in the culturing room of 30 DEG C ± 2 DEG C.Each process divides 3 groups of cultivations,
Often group 3 bottles, every bottle graft kind 10～15 pieces of ovules.
In order to ovule growth state is more preferable, BT culture medium is added heteroauxing and gibberellins.
Preferably, described BT culture medium adds heteroauxing and gibberellins；Described heteroauxing
Addition be 8-12 μm ol/L, the addition of described gibberellins is 4-6 μm ol/L.
1.3.2 the Growth trends of the ovule of BT culture medium culturing is observed
Observe DPC=5,10,15,20,25,30 and (represent the natural law after cultivating, DPC=0
I.e. Ovule development same day) Growth trends, it is found that when cultivating 30 days, the fiber growth of ovule
Reach stable maturity state.Therefore, the ovule cultivated 30 days is selected to measure the length of fiber.
While different culture media is cultivated, control medium is set, the most only adds corresponding content
Heteroauxing and the BT culture medium of gibberellins.Wherein, sucrose, potassium chloride and mannitol
Add before sterilizing；(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, brassinosteroid all use filtration sterilization, filtration sterilization
After add in the culture medium after sterilizing.
Material in each test group include palm fibre 1-61, palm fibre 263, RT white cotton fiber, green cotton CC28,
Green cotton G88's is several, and each process divides 3 groups of cultivations, often group 3 bottles, every bottle graft kind 10-15
The kinds of culture medium that piece ovule uses is as shown in test group 1-6.
Test group 1
Matched group: ovules culture medium is interpolation heteroauxing, gibberellins in BT culture medium,
The addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L；
Group 1: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Brassinosteroid, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of brassinosteroid is 0.05 μm ol/L；
Group 2: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Brassinosteroid, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of brassinosteroid is 0.1 μm ol/L；
Group 3: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Brassinosteroid, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of brassinosteroid is 0.5 μm ol/L；
Group 4: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Brassinosteroid, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of brassinosteroid is 1 μm ol/L.
Test group 2
Matched group: ovules culture medium is interpolation heteroauxing, gibberellins in BT culture medium,
The addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L；
Group 1: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.05 μm ol/L；
Group 2: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5 μm ol/L；
Group 3: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 5 μm ol/L；
Group 4: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate and brassinosteroid, the addition of heteroauxing is 10 μm ol/L, gibberellins
Addition be 5 μm ol/L, the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 5 μm ol/L, brassinosteroid
Content be 0.05 μm ol/L；
Group 5: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
(-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate and brassinosteroid, the addition of heteroauxing is 10 μm ol/L, gibberellins
Addition be 5 μm ol/L, the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 5 μm ol/L, brassinosteroid
Content be 0.2 μm ol/L.
Test group 3
Matched group: ovules culture medium is interpolation heteroauxing, gibberellins in BT culture medium,
The addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L；
Group 1: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Sucrose, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L,
The content of sucrose is 2g/L；
Group 2: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Sucrose, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L,
The content of sucrose is 5g/L；
Group 3: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Sucrose, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L,
The content of sucrose is 10g/L；
Group 4: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Sucrose, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L,
The content of sucrose is 15g/L.
Test group 4
Matched group: ovules culture medium is interpolation heteroauxing, gibberellins in BT culture medium,
The addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L；
Group 1: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Potassium chloride, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of potassium chloride is 0.01mol/L；
Group 2: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Potassium chloride, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of potassium chloride is 0.05mol/L；
Group 3: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Potassium chloride, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of potassium chloride is 0.08mol/L.
Test group 5
Matched group: ovules culture medium is interpolation heteroauxing, gibberellins in BT culture medium,
The addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5 μm ol/L；
Group 1: ovules culture medium be in BT culture medium interpolation heteroauxing, gibberellins,
Mannitol, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of mannitol is 3g/L；
Group 2: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Mannitol, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of mannitol is 5g/L；
Group 3: ovules culture medium: in BT culture medium add heteroauxing, gibberellins,
Mannitol, the addition of heteroauxing is 10 μm ol/L, and the addition of gibberellins is 5
μm ol/L, the content of mannitol is 10g/L.
Take 10 group training ovules of latter 30 days at random, take out after boiling water bath 5min, clear water
Rinse, carefully overlapping fibers separated with pin and make it stretch, measuring fibre length with ruler.
Wherein, the Cotton Gossypii of each kind takes 30 ovules the most at random and measures fibre length, is averaging
Value.Being analyzed the data of the fibre length obtained, the result obtained is as shown in table 2-5.
Rate of increase=(sample fibres length-matched group fibre length)/matched group fibre length * 100%.
The fibre length of each cotton variety in table 2 test group 1
During from table 2 it can be seen that the content of brassinosteroid is 0.05-1 μm ol/L, it is fine
Dimension length all has elongation, and wherein, the content of brassinosteroid is 0.1-0.5 μm ol/L, fiber
The rate of increase of length is higher.
The fibre length of each cotton variety in table 3 test group 2
During from table 3 it can be seen that the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.05-5 μm ol/L, it is fine
Dimension length all has elongation, and wherein, when being 5 μm ol/L with the content of (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, fiber is long
The rate of increase of degree is higher；Additionally, group 4 and group 5 are by (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate and brassinosteroid
Being used in combination, both play chemiluminescence, and the cotton fiber length obtained is longer；This
Outward, when (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5 μm ol/L, 1 μm ol/L, 3 μm ol/L, and in Brassica campestris L element
Ester with the use of, its fibre length also has greatly improved.
The fibre length of each cotton variety in table 4 test group 3
During from table 4, it can be seen that the content of sucrose is 2-15g/L, its fibre length all has
Elongation, wherein, the content of sucrose is 5g/L, and the rate of increase of fibre length is higher.
The fibre length of each cotton variety in table 5 test group 4
As can be seen from Table 5, when the content of potassium chloride is 0.01-0.08mol/L, its fiber
Length all has elongation, and wherein, the content of potassium chloride is 0.05mol/L, the increasing of fibre length
Long rate is higher.
The fibre length of each cotton variety in table 6 test group 5
As can be seen from Table 6, when the content of mannitol is 3-10g/L, its fibre length all has
Elongation, wherein, when the content of mannitol is 5g/L, the rate of increase of fibre length is higher.
The foregoing is only the preferred embodiments of the present invention, be not limited to the present invention,
For a person skilled in the art, the present invention can have various modifications and variations.All
Within the spirit and principles in the present invention, any modification, equivalent substitution and improvement etc. made,
Should be included within the scope of the present invention.
Claims (3)
1. the culture medium increasing cotton fiber length, it is characterised in that train at BT
Support and base adds second component, any one in following of described second component: jasmonic
Methyl ester, brassinosteroid and (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, sucrose, potassium chloride, mannitol；
When described second component is (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, and the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5-7
μmol/L；
When described second component is brassinosteroid and (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, described brassinosteroid
Content be 0.05-0.2 μm ol/L, the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is 0.5-5 μm ol/L；
When described second component is sucrose, and the content of described sucrose is 10-15g/L；
When described second component is potassium chloride, the content of described potassium chloride be 0.01mol/L or
0.08mol/L；
When described second component is mannitol, and the content of described mannitol is 3-10g/L；
Described BT culture medium is possibly together with heteroauxing and gibberellins；Adding of described heteroauxing
Dosage is 8-12 μm ol/L, and the addition of described gibberellins is 4-6 μm ol/L.
The culture medium of increase cotton fiber length the most according to claim 1, it is special
Levy and be, when described second component is (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate, and the content of described (-)-methyl cis-2-pent-2'-enyl-3-oxocyclopentylacetate is
0.5-5μmol/L。
3. the cultural method increasing cotton fiber length, it is characterised in that include following
Step: field Cotton Gossypii blooms and carries out selfing listing mark the previous day, takes Post flowering three days
The ovule of children's bell, then uses the increase cotton fiber described in any one of claim 1-2 long
The culture medium of degree carries out in vitro light culture；Cultivation temperature is 30 DEG C ± 2 DEG C.
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| CN100496225C (en) * | 2006-06-27 | 2009-06-10 | 中国农业科学院棉花研究所 | Cotton leafstalk tissue cultivation and high-differentiation rate material selective breeding method |
| BR122017020958B1 (en) * | 2007-05-08 | 2022-05-10 | Monsanto Technology Llc | Methods to improve the efficiency of cotton plant regeneration |
| CN100427589C (en) * | 2007-09-21 | 2008-10-22 | 河北农业大学 | Culture medium and culture method for cotton immature embryo in vitro culture and plantlet formation |
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| CN104109032A (en) | 2014-10-22 |
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