CN107980620A - A kind of the compound of plant chromosome doubles agent and method for doubling - Google Patents
A kind of the compound of plant chromosome doubles agent and method for doubling Download PDFInfo
- Publication number
- CN107980620A CN107980620A CN201711265252.7A CN201711265252A CN107980620A CN 107980620 A CN107980620 A CN 107980620A CN 201711265252 A CN201711265252 A CN 201711265252A CN 107980620 A CN107980620 A CN 107980620A
- Authority
- CN
- China
- Prior art keywords
- compound
- doubling
- agent
- plant
- double
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Classifications
-
- 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
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
- A01H1/08—Methods for producing changes in chromosome number
Landscapes
- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Life Sciences & Earth Sciences (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
A kind of reagent combination and method for doubling doubled using the composite drug progress plant chromosome without colchicine, the compound prescription prepared by using various kinds of cell mitotic inhibitor and Antitubulin is carried out plant chromosome and doubled, compared with existing chromosome doubling method, this method has the characteristics that less toxic, efficient, applied widely.Double agent, auxiliary agent and antidote three classes same-action compound be used cooperatively be the present invention main points and key, play the role of improve double effect and reduce drug induced injury.It can solve the abnormal existing Material growth for using colchicine damage after greatly, often causing processing the vegetable material of processing during plant chromosome doubles, deformity or even handle the death of material and double the problem of success rate is relatively low.It can more importantly solve the problems, such as that colchicine has mammal including humans hypertoxic and potential carcinogenesis, health and Environmental security to operating personnel to have very big harm.
Description
Technical field
Agent prescription and method for doubling are doubled the present invention relates to plant chromosome, belongs to phytobiology, plant biological skill
Art and field of plant breeding, it is particularly a kind of to obtain plant polyploid and monoploid is carried out doubling to become doubled haploid（Again
Claim dihaploid, pure and mild diploid）Method.
Background technology
Plant sexually revise again be phytobiology research, Plant Biotechnology and plant breeding important research content.
Compared with diploid, autopolyploid has the characteristics that some organ increases or metabolite content improve, for being sought with harvesting
Crop and asexually propagated crop for the purpose of foster organ have fabulous breeding utilization value；Artificial creation's polyploid can also be by open country
The inhereditary material of non-hibernating eggs and cultigen recombinates, and is bred as new crop；Doubling monoploids can obtain genotype homozygote plant pure lines simultaneously
Shorten the time limit of breeding；Odd-multiple polyploid（Such as triploid）Acquisition can be bred as aspermous watermelon, Zante currant, without seed citrus etc.
Without seed fruit, it can also cultivate and not produced seed bearing plant species, be primarily useful for the urban landscapings such as willow and speed
The cultivation of raw woods kind, is not produced the kind of willow catkins flying in the air；Can by different sibling species hybridization and then the allopolyploid obtained
For cultivating new species, new crop and being studied available for plant evolution.Therefore chromosome doubling have great theoretical research and
Commercialization value.These researchs all rely on a common technology --- and it is exactly chromosome doubling technology.Past frequently with dye
Doubling of chromosome technology is by the use of colchicine as doubling agent, but colchicine is difficult to meet the studies above to the effect that doubles of plant
Needs, main problem, which is colchicine, to be damaged the vegetable material of processing big, is often caused growth failure, deformity or even is located
Manage the death of material, in addition colchicine to double success rate relatively low.And colchicum is known as severe toxicity, to including humans
Mammal there is the toxicity of bigger and potential carcinogenesis, health and Environmental security to operating personnel have very big
Harm.Colchicine, high-efficiency low-toxicity plant chromosome method for doubling can be substituted because the invention is a kind of to be of great significance.
The content of the invention
The object of the present invention is to provide a kind of plant chromosome more less toxic than colchicine, efficient, applied widely
Method for doubling.
The method for doubling of plant chromosome provided by the present invention is to use various kinds of cell mitotic inhibitor and micro-pipe
The compound prescription that protein inhibitor is prepared carries out plant chromosome and doubles.
The active ingredient for doubling agent is following compound：Butralin（Chemical name：N- sec-butyl -4- tertiary butyl -2,
6- dinitroanilines；English name：Butralin；Molecular formula：C14H21N3O4；Molecular weight：295.3342）, amiprophos-methyl（Change
Scientific name claims：O- methyl-O- (2- nitro-4-methyls phenyl)-N- isopropylthio phosphonic amides；English name：AMIPROFOS
METHYL, AMP；Molecular formula：C11H17N2O4PS；Molecular weight：304.3）, oryzalin（Chemical name：3,5- dinitros-N', N'-
Dipropyl sulfanilamide (SN)；3,5- dinitros-N4, N4- propyl group sulfanilamide (SN)；English name：Oryzalin；Molecular formula： C12H18N4O6S；Molecule
Amount：346.3595）, nocodazole（Chemical name：Methyl-(5-- thenoyl -2- benzimidazolyls) carbamic acid；English name
Claim：Nocodazole；Molecular formula：C14H11N3O3S；Molecular weight：301.3204）, trefanocide（Chemical name：2,6- dinitro-N,
N- diη-propyl -4- 5-trifluoromethylanilines；English name：Trifluralin；Molecular formula：C13H16F3N3O4；Molecular weight：
335.28）, prodiamine（Chemical name：5- dipropylaminos-α, α, α -4,6- dinitros-o- toluidines；English name：
Procymidon；Molecular formula：C13H17N4O4；Molecular weight：350.3）, dithiopyr（Chemical name：S, S '-dimethyl -2- difluoros
Methyl -4- isobutyl group -6- trifluoromethyl pyridine -3,5- dithio formates；English name：Dithiopyr；Molecular formula：
C15H16F5NO2S2；Molecular weight：401.4）, diphenamide（Chemical name：N, N- dimethyl -2,2- acetyldiphenylamine；English
Title： DIPHENAMID；Molecular formula： C16H17NO；Molecular weight： 239.31）, pretilachlor（Chemical name：Chloro- N- (the 2,6- of 2-
Diethyl phenyl)-N- (2- Amongs) acetamide；English name：Pretilachlor；Molecular formula： C17H26ClNO2；Point
Son amount： 311.85）, pentyl xanthate（Chemical name：3,5- bis- chloro- N- (1,1- alkynyl dimethyls) benzamide；English name
Claim：Propyzamide；Molecular formula：C12H11Cl2NO；Molecular weight：256.13）, mefenacet（Chemical name：2- (1,3- benzene
And thiazol-2-yl oxygen)-N- methyl vinyls replace aniline；English name：Mefenacet；Molecular formula：C16H14N2O2S；Molecular weight：
298.3596）, isopropyl methoxalamine（Chemical name：Chloro- 2 ', the 6 '-diethyl-N- of 2- (2- Amongs) acetanil；English
Literary fame claims： Metolachlor；Molecular formula： C15H22ClNO2；Molecular weight：283.79）, chlorpropham（Chemical name：N-(3-
Chlorphenyl) carbamic acid isopropyl ester；English name：Chlorpropham；Molecular formula：C10H12ClNO2；Molecular weight：213.66）、
Pendimethalin（Chemical name：N- (1- ethyl propyls) -2,6- dinitro -3,4- dimethylanilines；English name：
Pendimethalin；Molecular formula：C13H19N3O4；Molecular weight：281.31）, Chem hoe（Chemical name：N- phenylcarbamic acids are different
Propyl ester；English name：Propham；Molecular formula：C10H13NO2；Molecular weight：179.22）；Above-claimed cpd is is made in the present invention
Double agent；
The medicament compound that doubles need to be with ethylene glycol（English name ethylene glycol, abbreviation EG）And/or dimethyl sulfoxide
（English name: Dimethyl sulfoxide）And/or toluene and/or dimethylbenzene are auxiliary agent, it acts as promote medicament dissolution
With infiltration of the reinforcement medicament in plant tissue and cell, medicament is promoted to reach the separate living tissues such as stem apex, bud point, the tip of a root and have
Growing point and inflorescence etc. can produce organ of multiplication and the position of offspring, to have the function that to double.Market can also be used to sell
Commercially available agricultural chemical auxiliary agent, including organosilicon auxiliary agent, adhesive agent and emulsifying agent, in the market it is common for TWEEN20, TWEEN60,
TWEEN80 etc., above-claimed cpd are the auxiliary agent used in the present invention；
The use for doubling to further include potassium phosphate and/or dipotassium hydrogen phosphate and/or potassium dihydrogen phosphate in processing, which rises slow
Solve foregoing agents toxicity, reduce processing material slow-growing, stagnation, exception, deformity and death etc. caused by medicament injures
Harmful effect, while the vegetable material to handle provides phosphorus potassium nutrition element necessary to growth, alleviation occurs after doubling processing
Scarce phosphorus and potassium deficiency symptom, such compound is used as antidote in the method.It can also be added in the antidote and contain it
In his the inorganic and nutrient solution of Organic nutrient, including MS nutrient solutions, Hoagland nutrient solutions and various mill water culture nutrient solutions
The change formula that various composition changes, this kind of occupation mode by for double the material after processing provide more fully comprehensive nutrition into
Point, but its nucleus is the potassium phosphate and/or dipotassium hydrogen phosphate and/or di(2-ethylhexyl)phosphate of 0.1%~2% mass volume percent content
Hydrogen potassium；
The treatment site doubled in processing further includes making for ABT root-inducing powders when being whole strain, root and the material for needing to transplant
With its concentration is 10~25ppm（Parts per million, parts per million）, it is the differentiation and growth for promoting root that it, which is acted on,
Plant survival rate after raising processing.Such compound（Medicine, reagent）Property be plant growth regulator.The growth
Conditioning agent is in addition to ABT root-inducing powders, or the auxins compound of other classes of accelerating agent containing plant growth regulator, including containing
There are heteroauxin, indolebutyric acid, methyl α-naphthyl acetate, the compound of compound sodium nitrophenolate class（Such as butyric acid potassium and/or Nafusaku）；
Composite drug is used as the main points of the present invention, foregoing four classes difference action compound（Double agent, auxiliary agent, antidote and life
Long conditioning agent）With the use of be the present invention core and key, can play improve doubling effect and reduce drug induced injury work
With.It is described to double agent and used in the form of being dissolved in water, butralin, amiprophos-methyl, oryzalin, nocodazole, trefanocide,
Prodiamine, dithiopyr, diphenamide, pretilachlor, pentyl xanthate, mefenacet, isopropyl methoxalamine, chlorpropham,
The ultimate density scope of the every kind of compound list product of Pendimethalin, Chem hoe is 0.01 μM~10mM；Ethylene glycol, dimethyl sulfoxide,
Toluene, the ultimate density scope of dimethylbenzene are 0.02~5%（V/V, volume percent content）；Potassium phosphate, dipotassium hydrogen phosphate, phosphorus
The ultimate density of acid dihydride potassium is 0.1%~2%（W/V, mass percentage content）；The concentration of ABT root-inducing powders and the like is
10~25ppm（Parts per million, parts per million）；Concentration used can for this in successive range any concentration and bag
Containing endpoints thereof；
It is described to double agent and be used in the form of coating agent for seed, bulking value hundred of every kind of compound list product in coating agent
It is 0.0001~1% to divide than content；
The medicament compound is commercially available unless otherwise specified, including can be from experiment consumptive material supplier, chemical industry
Factory, chemical reagent（Medicament）Obtained on company, pesticide manufacturer and its agency, pesticide company, various Agricultural Materials shops or agricultural materials market.
Different preparation ways includes analyzing pure level, chemical pure level and various various concentrations, the commercial preparation of different content；
It is described it is compound to double agent be not that foregoing all medicines must be appeared in same formula, because different agents are not to of the same race
The damaging action of class plant is of different sizes, and the action effect of unifacial leaf and dicotyledon is also differed, therefore in the application
Need overall balance to consider to double effect and survival rate, formula is combined for the different medicament of different Plants selections.Specifically,
The agent that doubles used in the present invention doubles effect and damage journey to unifacial leaf and dicotyledon, to perennial and annual plant
Degree need to take different compound combination and concentration combination there are significant difference according to different plant classifications；
The compound combination can be foregoing to double in agent, auxiliary agent, antidote and growth regulator any 2 kinds and of more than two kinds
All possible combinations of whole；
Workable plant scope of the present invention is all plants for carrying out mitosis and meiosis, for doubling to locate
The processed material of reason includes the intact plant of these plants, a part for plant, organ, tissue and cell etc.；This method is repaiied
Change the chromosome doubling that can also be used for the animal and fungi with mitosis and meiosis ability.These plant scopes include
But it is not limited to：Watermelon, cucumber, muskmelon, capsicum, eggplant, tomato, potato, arabidopsis, sweet potato, citrus, shaddock, sugarcane, rubber
Tree, banana, mango, pseudo-ginseng, corn, rice, wheat, cotton, soybean, rape, wild cabbage, Chinese cabbage, grape, apple, pears, pawpaw,
Peanut, sorghum, tobacco, clover, cherry, willow, willow, cassava, oil palm, barley, oat, sunflower, millet, beet, lotus, hundred
Conjunction, peach, Lee, apricot, chrysanthemum, tea tree, pepper, pumpkin, wax gourd, sponge gourd, balsam pear, Kidney bean, cowpea, broad bean, chick-pea, lupin,
Clover, ginseng, radix glycyrrhizae, English walnut, Kiwi berry, lichee, longan, jujube, pseudo-ginseng, persimmon, cauliflower, wild cabbage, spinach, strawberry,
Blueberry, mulberry tree, Chinese rose, orchid, rose, iris, gladiolus, China pink, barley, tree peony, Chinese herbaceous peony, Chinese yew, ginkgo, lettuce,
Leek, green onion, round onions, plum blossom, dandelion；The application range of the present invention further includes the chromosome doubling to various ploidy difference plants
And double the acquisition of rear material, containing aneuploid, haploid,diploid,triploid,tetraploid, pentaploid, hexaploid, seven times
Body, octoploid, nonuploid, decaploid, the dodecaploid, ten hexaploids.
The processing method of the present invention that doubles is included to treatment site, processing time and processing material and processing mode etc.
The selection and combination of factor：
Treatment site：Usually using plant shoot apical meristem, floral meristem and rataria in growth course to be main right
As can also make medicament reach growing point by the processing to the non-separate living tissue such as root, stem and leaf using absorbability and conductibility effect；
Processing time：Day differed from 1min to 50, Best Times are different because of floristics, the material of processing and processing method；
Processing mode：By the various modes of action directly or indirectly contacted, make chromosome doubling and produce fertile offspring；
A. non-germination seed is handled：Can take seed soaking, seed pelleting, spray, smear, culture medium addition, blotting paper and all kinds of right
Liquid has the modes such as the material parcel of adsorptivity, containment to carry out doubling to handle；
B. handle and sprouted（Germination）Seed or/and seedling：It can use and the insecticide-applying way such as impregnate, smear, spraying, being added dropwise, handle
When can take the method that growing point is exposed as needed, including divest/cut off peripheral blade, leaf sheath, cotyledon, spire etc.
Organ and tissue, effect is doubled to improve；Also medicament can be directly injected into growing point annex by the way of micro-injection；
C. the processing of root：Medicament dipping, water planting, sand culture, soil addition, artificial substratum addition, culture medium addition, spray can be used
The mode such as apply, processing procedure can be continuous processing, or batch process or short-term discontinuity treatment or/and disposable processing；
D. tender growing point, bud point, stem apex and inflorescence tissue treatment：Drop, coating can be used（Smear）Or with containing solution
Absorbent cotton, filter paper and all kinds of materials with adsorption capacity and the administration such as pending material method, during use
Further include in the medicament to the present invention and add thickener, tackifier etc., to strengthen the adhesive ability of medicine；Micro note can also be used
The mode penetrated, medicament is directly injected near growing point or/and organ of multiplication；
E. the processing of immature seed and rataria：Can use impregnate, smear, spraying, drop, injection, coating, culture medium add,
The modes such as pollen tube imports, filigree imports, floral organ imports achieve the purpose that to double.
The positive effect of the present invention：Compared with the method for doubling using colchicine, medicament compound used in this method
Concentration is low, dosage is few, has the characteristics that low toxicity, low-residual, degradable and environmental pollution are small, especially to people and other animals
Low toxicity, the field death rate of plant is low after processing, plant strain growth is abnormal incidence is low, it is higher to double rate, is a kind of effective
Chromosome doubling method.
Embodiment
The method of following offers is the compound medicine for being illustrated the effect of the method for the present invention with example rather than being used this method
Agent species, concentration, combination and use scope are limited or/and limited.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples, is commercially available unless otherwise specified, including can
From experiment consumptive material supplier, chemical plant, chemical reagent（Medicament）Company, pesticide manufacturer and its agency, pesticide company, the various means of agricultural productions
It is commercially available on shop or agricultural materials market.
The molar concentration of various reagents solution is with the active constituent content of the compound of actual purchase in following embodiments
Obtained from being prepared after being computed.
The chromosome doubling of one monocotyledon rye of embodiment
First, preparation of reagents
1. double the preparation of agent mother liquor
It is foregoing double agent need it is every kind of individually be configured to mother liquor, the used time, every kind of mother liquor was as follows by recipe requirements dilution
Unified to prepare, the preparation method of reagent/medicament is the routine techniques used in common lab：
50ml dimethyl sulfoxides are measured into the volumetric flask of 1000ml, by final volume be 1000ml, ultimate density is 10mM and root
The solid medicine for calculating and weighing respective numbers according to the effective content of every kind of medicine is added in dimethyl sulfoxide, is slowly stirred to complete
Dissolving；The ethylene glycol of 200ml is added, is stirred well to and is uniformly dissolved；Add distilled water and be settled to 1000ml, be sufficiently stirred mixing
Afterwards, brown bottle is loaded, and it is spare to be put into 4 degrees Celsius of refrigerators；
2. the compound preparation for doubling agent
Distilled water of the 500ml by sterilizing is added in the volumetric flask of 1000ml, it is female to draw above-mentioned reagent respectively with pipettor
Liquid is added in distilled water, the amount of absorption is 3 ml of methyl amine grass phosphine, 3 ml of diphenamide, 3 ml of butralin, 1 ml of trefanocide,
1 ml of pretilachlor, ammonia sulphur 1 ml of spirit, are then settled to 1000ml with distilled water.Each component ultimate density in the solution point above
Wei not 30 μM of methyl amine grass phosphine, 30 μM of diphenamide, 30 μM of butralin, 10 μM of trefanocide, 10 μM of pretilachlor, 10 μ of ammonia sulphur spirit
M。
2nd, the preparation and pre-treatment of vegetable material to be doubled --- seed treatment and germination
The rye seed of germinating capacity is taken, at room temperature（18~25 DEG C or so）With clear water soaked overnight（14~18 it is small when）, make
Seed imbibition；Then rye seed is put into the germination box for being covered with germination paper or filter paper, phjytotron at 20~25 DEG C,
Temperature automatically controlled incubator or at room temperature moisturizing culture, are periodically rinsed 3 times with clear water in the morning, afternoon and evening daily, until root long reach 0.5~
1.0 cm。
3rd, double to handle
The above-mentioned rye seed to have germinateed is put into the beaker of 200ml, doubles agent, it is desirable to which medicament must not have children completely
Bud and young root, continue to be put into constant temperature or room temperature（18~25 DEG C）Under the conditions of cultivate, the time for 6~8 it is small when.Then cleaned with clear water
And continue culture 8~24 it is small when.
4th, effect detection is doubled --- cytology detects
The tip of a root is taken to double effect using interim pressed disc method observation cell chromosome number, inspection under an optical microscope.Wherein not
Rye diploid root-tip cells double, normal only have 14 chromosomes, and double successful tetraploid root-tip cells tool
There are 28 chromosomes.
5th, effect is doubled
100 young shoot tips of a root doubled after processing are taken to carry out doubling validity check, the rate of doubling reaches 100%；Colchicine doubles
Effect reaches 100%.This method doubles effect without significant difference with colchicine to the rye tip of a root.
The chromosome doubling of two dicotyledon pea of embodiment
First, preparation of reagents
1. double the preparation of agent mother liquor
It is identical with example one
2. the compound preparation for doubling agent
Distilled water of the 500ml by sterilizing is added in the volumetric flask of 1000ml, it is female to draw above-mentioned reagent respectively with pipettor
Liquid is added in distilled water, the amount of absorption is 5 ml of butralin, 3 ml of methyl amine grass phosphine, 3 ml of prodiamine, 1 ml of pentyl xanthate,
1 ml of mefenacet, 1 ml of dithiopyr, are then settled to 1000ml with distilled water.More than each component in the solution final
Concentration be respectively 50 μM of butralin, 30 μM of methyl amine grass phosphine, 30 μM of prodiamine, 10 μM of pentyl xanthate, 10 μM of mefenacet,
10 μM of dithiopyr.
2nd, the preparation and pre-treatment of vegetable material to be doubled --- seed treatment and germination
The pea seed of germinating capacity is taken, at room temperature（18~25 DEG C or so）With clear water soaked overnight（14~18 it is small when）, make
Seed imbibition；Then pea seed is put into the germination box for being covered with germination paper or filter paper, phjytotron at 20 degrees Celsius,
Temperature automatically controlled incubator moisturizing culture, daily in the morning, afternoon and evening periodically with clear water rinse 3 times, until radicle dash forward break in the seed coat, root long reaches
0.5~1.0 cm, this process generally require 2 days.
3rd, double to handle
The above-mentioned pea seed to have germinateed is put into the beaker of 200ml, add in this example prepare compound double agent, it is desirable to medicine
Agent must not there be the pea and young root of germination completely, continue to be put into constant temperature or room temperature（18~25 DEG C）Under the conditions of cultivate, the time 6
~8 it is small when.Then cleaned up with clear water, then put back to germination box in continue culture 48~72 it is small when.
4th, effect detection is doubled --- morphology and cytology detection
By after doubling the pea seedlings of processing and taking out, it is possible to find obvious metamorphosis, prominent features are that bud and root development shorten
Thicker, especially it is short and thick to expand swelling, form for the tip of a root；Take the tip of a root under an optical microscope using interim pressed disc method observation cell dye
Colour solid number, inspection double effect.Pea diploid root-tip cells do not double, normal only have 7 pairs（14）Chromosome,
And it is even more with 28 chromosomes to double successful root-tip cells.
5th, effect is doubled
100 young shoot tips of a root doubled after processing are taken to carry out doubling validity check, the rate of doubling reaches 100%；
Embodiment one and embodiment two are only to test compound to double action effect of the agent to monocotyledon and dicotyledon, institute
Not carry out Detoxified treatment and later stage seedling culture and transplanting；If needing later stage culture seedling and transplanting, should carry out Detoxified treatment and
Rooting treatment.
But present method be advantageous in that：
1. processing time is short, when colchicine needs processing 26 small or so, this method only need 6 it is small when or so, improve drug concentration
It may be further improved with treatment temperature and double effect and shorten processing time；
2. double medicament compound used in this method without severe toxicity, and colchicine is severe toxicity, thus health to operating personnel and
Harm to environment is much smaller than the former processing method using colchicine to double agent.
The chromosome doubling of three corn monoploid rataria of embodiment
First, preparation of reagents
1. reagent mother liquor
It is identical with example one；
2. the compound preparation for doubling agent
Distilled water of the 60ml by sterilizing is added in the volumetric flask of 100ml, draws above-mentioned reagent mother liquor respectively with pipettor
Add in distilled water, the amount of absorption is 0.5 ml of methyl amine grass phosphine, 0.5 ml of nocodazole, 0.3 ml of Chem hoe, chlorpropham
0.3 ml, 0.1 ml of pentyl xanthate, 0.1 ml of Pendimethalin, 0.1 ml of isopropyl methoxalamine, are then settled to distilled water
100ml.The ultimate density of each component in the solution is respectively 50 μM of methyl amine grass phosphine, 50 μM of nocodazole, 30 μ of Chem hoe above
M, 30 μM of chlorpropham, 10 μM of pentyl xanthate, 10 μM of Pendimethalin, 10 μM of isopropyl methoxalamine.
3. the preparation of antidote：By 0.2g potassium dihydrogen phosphates add 100ml distilled water in be configured to 0.2% it is water-soluble
Liquid, then adds the urea of 0.5g；
4. the preparation of ABT rooting powder solutions：ABT root-inducing powders 6 are used in this example, it is limited by the good woods science and technology of Beijing middle forest
Company produces, and it is 20ppm to be made into concentration by the specification requirement provided on packing box（Parts per million, Bai Wanfen
Rate）Aqueous solution；
2nd, the preparation and pre-treatment of vegetable material to be doubled
1. haploid induction
This experiment rataria used is using normal diploid corn as female parent, with single times with maternal haploid induction ability
Body induced material is male parent, is obtained by the method for artificial hybridization., need to be to maternal female fringe set before maternal female fringe filigree is grown
Bag is to prevent pollen contamination；After maternal female fringe filigree is grown, give maternal female fringe to pollinate with paternal pollen, obtain two times of heterozygosis
Body seed and monoploid seed.Controlled pollination and prevent the method that foreign pollen pollutes for generally use in maize genetic and breeding
Ordinary maize pollination technique.This pollinating method can produce 1.5%~23% monoploid seed, its ratio is not because of induction
It is same and different, remaining mainly diploid heterozygosis seed.Female parent can be cenospecies, local varieties, synthetic variety, self-mating system and
Artificial synthesized colony etc. or other Zea materials of energy self-reproduction, no matter whether it is in hereditary homozygotic state；
Male parent induced material can be haploid inducing line such as Stock 6, agricultural university's height lure No. 1, RWS, KEMS, KMS, ZMS,
CAUHI1, CAUHI5 etc. or by not isogeneous induction system prepare various forms of cenospecies, including the seed of single cross, the seed of double cross,
Triple hybrid, collective species etc., and its progeny material of different segregating generations.This experiment haploid inducing line used is that agricultural university's height lures
No. 1 (public can obtain from China Agricultural University, recorded the non-patent literature of the material and be：Liu Zhizeng, Song with bright 2000,
The selection and breeding and identification of Haploid Inducer with High Frequ ency Parthenogenesis in Maize, Acta Agronomica Sinica, 26 (5)：570 1 574)；
2. the acquisition of rataria
5~30 days after pollination, maternal immature fruit ear is removed, removes bract and filigree, fruit ear surface is by 20min's
After hypochlorous acid or dioxygen water sterilization, and by manually taking out rataria with sterilized scalpel.The rataria of taking-up, which is put into, to be sterilized
Pure water in of short duration preservation, after the rataria of acquisition reaches certain amount, carry out doubling to handle together；
3rd, double to handle
By the rataria of taking-up it is drained after, be put into culture dish, add it is foregoing doubles agent to do not cross rataria, processing time 20
~30min；Then outwell liquid and rinsed repeatedly with sterile water at least 3 times, wash away the liquid of remnants；
4th, double to post-process
Detoxified treatment：Rataria is put into 0.2%（W/V）Potassium dihydrogen phosphate in, soak 20min；
IMMATURE EMBRYOS CULTURE：It uniformly will neatly put on the culture dish containing full nutrient medium, note by the rataria for doubling processing
Anticipate embryo down, scultellum upward, light culture 6~10 days at 26 DEG C（Generally 7 days）, then it is transferred to illumination cultivation 2~3 weeks,
Continue to cultivate bud and root until plantlet grows health, become the seedling with viability.In the process, according to color
Rataria or seedling are divided into diploid and monoploid by the presence or absence of mark, and purpuric is diploid, and the possibility of no purple is single
Times body.
5th, sprigging
When transplanting seedlings, seedling can be divided into monoploid and diploid again by mark properties such as colors, the two will be moved respectively
Plant.When using ABT rooting powder solutions leaching root processing 2 small before transplanting；Then transplant seedlings in the culturing pot equipped with Nutrition Soil into
Row hardening adapts to, and is transplanted to again in greenhouse or open country after a week.Plant after transplanting presses normal cultivation management, bagging before blooming,
Post flowering selfing is until harvest arrives ripe seed.
6th, cytology detects
A tip of a root detection cell chromosome number can be taken when young root is grown and reaches 1~2cm long, inspection doubles effect.Wherein wrap
Containing doubling successful liploid plant（Cell）With 20 chromosomes, successful haplobiont is not doubled（Cell）Have
10 chromosomes, tetraploid plant（Cell）With 40 chromosomes.
7th, field is verified
Plant loose powder rate and the solid strain rate in the maturity period statistics in corn florescence
Seedling is monoploid or diploid by the rataria that color mark distinguishes before processing, and monoploid is all transplanted, and diploid is only
50 plants of transplanting.By above-mentioned sprigging into greenhouse.Carried out after slow seedling by the normal management measure of greenhouse production, florescence statistics
Monoploid loose powder strain rate and pollination self, after plant maturation again by seed on fruit ear whether there is color separated distinguish monoploid and
Diploid, counts the setting percentage of haplobiont.Gained seed progeny takes the planting patterns of head progeny row, passes through form between strain in row
The presence or absence of neat degree and seed purple mark of character and separation situation judge haploid to double effect.Result of the test table
It is bright：The method doubling monoploids rate (loose powder rate) of the present invention is up to 44.4%~64.7%, and average out to 51.9% is (because material difference has
Institute's difference)；Setting percentage after doubling is 36.4%~62.5%, and (because of material difference, difference, notebook data are average out to 48.1%
The result of bench-scale testing).It is tetraploid plant to find 2 plants in the offspring of liploid plant, and it is preferable to show that this method has
Chromosome doubling effect.
One IMMATURE EMBRYOS CULTURE of table doubles result of the test
| Fruit ear is numbered | Rataria number | Haploid embryo number | Number of seedling | Nursery transplant number | Loose powder strain number | Solid strain number |
| 1 | 121 | 19 | 17 | 17 | 11 | 4 |
| 2 | 136 | 21 | 18 | 18 | 8 | 4 |
| 3 | 128 | 18 | 18 | 17 | 8 | 5 |
| It is total | 385 | 58 | 53 | 52 | 27 | 13 |
| Ratio | 15.06% | 91.3% | 98.1% | 51.9% | 48.1% |
In actual application, this method can it is modified optimization and make more efficient., can be according to rataria face when obtaining rataria
Color distinguishes monoploid and diploid, and it is diploid to have the rataria that purple marks, and the rataria of no purple mark may be monoploid, only
Leave the rataria of no purple mark；During IMMATURE EMBRYOS CULTURE, the rataria for showing purple mark or plant are constantly eliminated, can be improved
The efficiency that monoploid differentiates and doubles.
Example IV corn haploid plantlets chromosome doubling
First, preparation of reagents
1. reagent mother liquor
It is identical with example one.
2. the compound preparation for doubling agent
10L tap water, well water, river water, water in a canal or other clear water are added in the agricultural atomizer of 15L, are measured respectively with graduated cylinder
Above-mentioned reagent mother liquor is added in sprayer, and the amount of addition is respectively 75 ml of methyl amine grass phosphine, 75 ml of nocodazole, oryzalin
45 ml, 45 ml of chlorpropham, trefanocide 30ml, 15 ml of pentyl xanthate, 15 ml of isopropyl methoxalamine, 15 ml of Pendimethalin,
The dimethyl sulfoxide of 50ml is added, 15L graduation marks is then added water to and is uniformly mixed.More than each component in the solution most
Final concentration is respectively 50 μM of methyl amine grass phosphine, 50 μM of nocodazole, 30 μM of oryzalin, 30 μM of chlorpropham, 20 μM of trefanocide, penta
10 μM of alkynes grass amine, 10 μM of isopropyl methoxalamine, 10 μM of Pendimethalin.
3. the preparation of antidote：It is identical with example three；
4. the preparation of ABT rooting powder solutions：It is identical with example three；
2nd, the preparation and pre-treatment of vegetable material to be doubled
Haploid preparation method
1. haploid induction
It is identical with embodiment three
2. monoploid seed is selected
After seed development maturation, female fringe is harvested.Monoploid and diploid are distinguished by the color mark on seed on fruit ear, is had
The seed of purple mark is diploid, and the seed of no purple mark may be monoploid, only retain monoploid seed for next
The experiment of step；
3. obtained monoploid seed is planted identical with conventional corn to big Tanaka, field management；
3rd, double to handle and double to post-process
1. when the monoploid seed sprouting sowed and when seedling grows to the 1 core phase of 1 leaf, with spray before the daily dusk sunset of fine day
Day with fog uniformly sprays downwards from seedling top, it is desirable to be especially covered with plant in corncob liquid and along seedling to flow down
Drop down；
2. spray doubles the next morning of agent, with atomizers spray antidote and root-inducing powder；
3. hereafter spray every night doubles agent, antidote and root-inducing powder are sprayed morning next day, repeat this operation until seedling is grown to
6 leaf phases terminated；
4th, the field verification of effect is doubled
Loose powder rate and the solid strain rate in the maturity period statistics of corn florescence plant
After doubling processing, plant is normally carried out field management, florescence statistics monoploid loose powder strain rate and pollination self, plant into
The setting percentage of statistics haplobiont after ripe.Result of the test shows：The method doubling monoploids rate (loose powder rate) of the present invention is reachable
60.2%, average out to 58.8% (the difference because of material difference)；Setting percentage after doubling is up to 45.4%, average out to 43.4%
(the difference because of material difference).Compared with using the method for doubling of colchicine, medicament compound is to ring used in this method
The pollution in border is small, less toxic, low-residual, degradable, and the field death rate after processing is low, plant strain growth abnormality incidence is low, operation
Simply, double that rate is higher, be a kind of effective ways of chromosome doubling.
Two seedling of table doubles result of the test
| Repeat | Handle strain number | Survive strain number | Loose powder strain number | Solid strain number | Survival rate（%） | Loose powder rate（%） | Setting percentage（%） |
| 1 | 260 | 201 | 153 | 115 | 77.3 | 58.8 | 44.2 |
| 2 | 260 | 207 | 149 | 118 | 79.6 | 57.3 | 45.4 |
| 3 | 249 | 199 | 150 | 101 | 79.9 | 60.2 | 40.6 |
| It is total | 769 | 608 | 452 | 334 | 79.1 | 58.8 | 43.4 |
Antidote used also is added in other inorganic and organic nutrient solution for cultivating and is used cooperatively in the present invention, including everybody is ripe
The change formula that the various composition of MS nutrient solutions, Hoagland nutrient solutions and the various mill water culture nutrient solutions known changes, but its core
Component is 0.1%~2%（W/V, mass percentage content）Potassium phosphate and/or dipotassium hydrogen phosphate and/or potassium dihydrogen phosphate.
This method effectively can double plant chromosome, but this method still have the potentiality of improvement and optimization for
More floristics outside embodiment, improvement and optimization to this method will likely further improve it and double effect；To we
The modification of method can also be used for the chromosome doubling of the animal and fungi with mitosis and meiosis ability；And to ability
It is any in domain to be familiar with for the personnel of the technical principle, can be according to above-mentioned after the inspiration of content of the present invention is subject to
Illustrate to be improved or convert, and all these modifications and variations should all belong to the protection domain of appended claims of the present invention.
Claims (9)
1. a kind of the compound of plant chromosome doubles agent, it is characterized in that：（1）It is compound to double agent as various kinds of cell mitosis suppression
Agent and the compound prescription of Antitubulin composition, doubling agent includes following compound：Butralin, amiprophos-methyl, peace sulphur
It is spirit, nocodazole, trefanocide, prodiamine, dithiopyr, diphenamide, pretilachlor, pentyl xanthate, mefenacet, different
Methoxalamine, chlorpropham, Pendimethalin, Chem hoe, these solvent versions for doubling medicament to be dissolved in water use, often
A kind of ultimate density scope for doubling agent list product is 0.01 μM~10mM；（2）Potassium phosphate, phosphoric acid hydrogen two are further included in compound prescription
Potassium and potassium dihydrogen phosphate are at least one, using it as antidote, are represented with mass percentage content, its final concentration range is
0.1%~2%；（3）It is at least one that ethylene glycol, dimethyl sulfoxide, toluene and dimethylbenzene are further included in compound prescription, as auxiliary agent, is pressed
Volume percent content represents that the ultimate density scope of each auxiliary agent is 0.02~5%；（4）ABT is further included in compound prescription
Root-inducing powder or its analog, as growth regulator, its final content is 10~25ppm；The compound formula for doubling agent includes upper
State all possible combinations of any two above compound.
2. compound as described in claim 1 doubles agent, it is characterized in that：It is compound to double agent and used in the form of coating agent for seed,
Weight percent content of every kind of compound list product in coating agent is 0.0001~1%.
3. compound as described in claim 1 doubles agent, it is characterized in that：Added in antidote it is various it is inorganic with organic nutrition into
Divide, include the change formula that the various composition of MS nutrient solutions, Hoagland nutrient solutions and various mill water culture nutrient solutions changes, but its core
Heart component is containing at least one of 0.1%~2% potassium phosphate, dipotassium hydrogen phosphate and potassium dihydrogen phosphate component.
4. compound as described in claim 1 doubles agent, it is characterized in that：Growth regulator is alternatively other plant auxins
Compound, including containing heteroauxin, indolebutyric acid, methyl α-naphthyl acetate, compound sodium nitrophenolate class compound, such as butyric acid potassium, methyl α-naphthyl acetate
Sodium is at least one.
A kind of 5. compound method for doubling agent progress plant chromosome and doubling of usage right requirement 1, it is characterized in that main step
It is rapid as follows：（1）The preparation of reagent,（2）The preparation and pre-treatment of vegetable material to be doubled,（3）Double to handle,（4）Locate after doubling
Reason,（5）Double effect detection and verification.
6. the method that the plant chromosome as described in claim 5 doubles, it is characterized in that：Double to handle workable plant scope
Including all plants for carrying out mitosis and meiosis, it, which doubles the processed material of processing, includes the complete plant of these plants
Strain, a part for plant, organ, tissue and cell；It can also be used for having the animal of mitosis and meiosis ability and true
The chromosome doubling of bacterium.
7. the method that the plant chromosome as described in claim 5 doubles, it is characterized in that：Processing time is doubled from 1min to 50
My god.
8. the method that the plant chromosome as described in claim 5 doubles, it is characterized in that：Doubling processing mode is included by various
The directly mode of action of contact and mediate contact makes chromosome doubling, specific as follows：
A. non-chitting piece is handled：Take seed soaking, at least one mode of seed pelleting carries out doubling to handle；
B. it is at least one that chitting piece, seedling are handled：Can use the insecticide-applying way that impregnates, smear, spraying, during processing according to
Need that the method that growing point is exposed can be taken, including divest/cut off peripheral blade, leaf sheath, cotyledon, spire organ and group
Knit, effect is doubled to improve；Medicament can be also directly injected near growing point by the way of micro-injection；
C. the processing of root：Medicament dipping, water planting, soil addition, artificial substratum addition, culture medium addition manner, processing can be used
Process can be continuous processing, or batch process or short-term discontinuity treatment or disposable processing；
D. tender growing point, bud point, stem apex and inflorescence tissue treatment：Drop, coating can be used or with the degreasing containing solution
Cotton, filter paper and all kinds of materials with adsorption capacity carry out packaging method, are further included during use and double agent addition to compound
Thickener, tackifier, to strengthen the adhesive ability of medicine；Also medicament can be directly injected into life by the way of micro-injection
Near long point；
E. the processing of immature seed and rataria：Can use impregnate, smear, spraying, drop, injection, coating, culture medium add
Mode achievees the purpose that to double.
9. the application of the plant chromosome method for doubling as described in claim 5, it is characterized in that：Application range includes various ploidies
The chromosome doubling of plant, containing aneuploid, haploid,diploid,triploid,tetraploid, pentaploid, hexaploid, heptaploid,
Octoploid, nonuploid, decaploid, the dodecaploid, ten hexaploids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711265252.7A CN107980620B (en) | 2017-12-05 | 2017-12-05 | Composition for chromosome doubling of corn haploid seedlings |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711265252.7A CN107980620B (en) | 2017-12-05 | 2017-12-05 | Composition for chromosome doubling of corn haploid seedlings |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN107980620A true CN107980620A (en) | 2018-05-04 |
| CN107980620B CN107980620B (en) | 2021-03-16 |
Family
ID=62035572
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711265252.7A Active CN107980620B (en) | 2017-12-05 | 2017-12-05 | Composition for chromosome doubling of corn haploid seedlings |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107980620B (en) |
Cited By (14)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108935086A (en) * | 2018-08-14 | 2018-12-07 | 吉林农业科技学院 | A kind of chemical seed-soaking liquid and preparation method thereof for corn doubling monoploids |
| CN109122293A (en) * | 2018-08-29 | 2019-01-04 | 广西壮族自治区农业科学院玉米研究所 | It is a kind of to double the haploid method of corn using herbicide |
| CN110192525A (en) * | 2019-06-28 | 2019-09-03 | 大连大学 | A kind of cultural method of blueberry polyploid |
| WO2019178680A1 (en) * | 2018-03-21 | 2019-09-26 | Canopy Growth Corporation | Methods for inducing polyploidy in cannabis |
| CN110732025A (en) * | 2018-07-19 | 2020-01-31 | 中国人民解放军军事科学院军事医学研究院 | Animal model of cell-in-cell structures and preparation method thereof |
| WO2021133922A1 (en) * | 2019-12-23 | 2021-07-01 | Calyxt, Inc. | Modified ploidy levels of a cannabis plant |
| CN113170727A (en) * | 2021-04-27 | 2021-07-27 | 淮阴工学院 | Breeding method of catkin willow plants |
| CN113383705A (en) * | 2021-07-19 | 2021-09-14 | 江西省农业科学院园艺研究所 | Method for breeding polyploid kiwi fruit |
| CN114041463A (en) * | 2021-11-09 | 2022-02-15 | 昆明学院 | Agents and method for reduplicating and reducing damage and improving mutagenesis rate of gynura bicolor chromosome |
| CN114521489A (en) * | 2022-03-04 | 2022-05-24 | 南通大学 | Method for inducing willow chromosome doubling at colchicine variable temperature |
| CN115500259A (en) * | 2022-10-28 | 2022-12-23 | 绍兴市农业科学研究院 | Method for overcoming pollenless of progeny of interspecific cross of pumpkin |
| CN115669535A (en) * | 2022-10-11 | 2023-02-03 | 中国林业科学研究院经济林研究所 | Method for efficiently inducing persimmon polyploidy by treating seeds with colchicine |
| CN117502225A (en) * | 2023-11-28 | 2024-02-06 | 中国热带农业科学院热带作物品种资源研究所 | Induction method and application of capsicum |
| CN117530173A (en) * | 2023-12-27 | 2024-02-09 | 中国热带农业科学院热带作物品种资源研究所 | Method for creating new germplasm of pepper tetraploid by utilizing stem tip induction |
Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631101A (en) * | 2003-12-22 | 2005-06-29 | 刘文革 | Excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique |
| WO2007038075A1 (en) * | 2005-09-21 | 2007-04-05 | Pioneer Hi-Bred International, Inc. | Doubling of chromosomes in haploid embryos |
| CN103053413A (en) * | 2012-12-16 | 2013-04-24 | 四川农业大学 | Chemical corn double haploid young embryo processing method |
| CN103053414A (en) * | 2011-10-24 | 2013-04-24 | 中国农业大学 | Method for doubling corn haploid by herbicide and special herbicide of method |
| CN103929950A (en) * | 2011-07-20 | 2014-07-16 | 凯伊玛生物农业科技有限公司 | Maize plants having a partially or fully multiplied genome and uses thereof |
| CN105340538A (en) * | 2015-11-11 | 2016-02-24 | 中国农业科学院棉花研究所 | Chromosome doubling method for cotton growing points |
| CN105940812A (en) * | 2016-05-31 | 2016-09-21 | 湖南生物机电职业技术学院 | Ploidy mutagenesis method of brier grape seeds dipped by propyzamide |
| CN106035068A (en) * | 2016-06-23 | 2016-10-26 | 成都市农林科学院 | Method for breeding leaf mustard type rape varieties and materials by rape double haploid inducing line |
-
2017
- 2017-12-05 CN CN201711265252.7A patent/CN107980620B/en active Active
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1631101A (en) * | 2003-12-22 | 2005-06-29 | 刘文革 | Excised mutagenesis tetraploid method of water melon and ploidy early stage certification technique |
| WO2007038075A1 (en) * | 2005-09-21 | 2007-04-05 | Pioneer Hi-Bred International, Inc. | Doubling of chromosomes in haploid embryos |
| CN103929950A (en) * | 2011-07-20 | 2014-07-16 | 凯伊玛生物农业科技有限公司 | Maize plants having a partially or fully multiplied genome and uses thereof |
| CN103053414A (en) * | 2011-10-24 | 2013-04-24 | 中国农业大学 | Method for doubling corn haploid by herbicide and special herbicide of method |
| CN103053413A (en) * | 2012-12-16 | 2013-04-24 | 四川农业大学 | Chemical corn double haploid young embryo processing method |
| CN105340538A (en) * | 2015-11-11 | 2016-02-24 | 中国农业科学院棉花研究所 | Chromosome doubling method for cotton growing points |
| CN105940812A (en) * | 2016-05-31 | 2016-09-21 | 湖南生物机电职业技术学院 | Ploidy mutagenesis method of brier grape seeds dipped by propyzamide |
| CN106035068A (en) * | 2016-06-23 | 2016-10-26 | 成都市农林科学院 | Method for breeding leaf mustard type rape varieties and materials by rape double haploid inducing line |
Non-Patent Citations (6)
| Title |
|---|
| 武婷婷: "大豆除草剂药害及其有效缓解剂的筛选", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 王兆振等: "除草剂对作物的药害研究", 《农药科学与管理》 * |
| 赵等: "除草剂在植物离体染色体加倍上的应用 ", 《长江蔬菜》 * |
| 邵果园等: "氨磺乐灵在园艺植物倍性育种上的应用", 《中国南方果树》 * |
| 陶抵辉等: "生物多倍体诱导方法研究进展", 《生命科学研究》 * |
| 黄权军: "化学药剂诱导植物多倍体研究进展 ", 《安徽农学通报(上半月刊)》 * |
Cited By (16)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2019178680A1 (en) * | 2018-03-21 | 2019-09-26 | Canopy Growth Corporation | Methods for inducing polyploidy in cannabis |
| CN110732025A (en) * | 2018-07-19 | 2020-01-31 | 中国人民解放军军事科学院军事医学研究院 | Animal model of cell-in-cell structures and preparation method thereof |
| CN108935086A (en) * | 2018-08-14 | 2018-12-07 | 吉林农业科技学院 | A kind of chemical seed-soaking liquid and preparation method thereof for corn doubling monoploids |
| CN109122293B (en) * | 2018-08-29 | 2022-04-05 | 广西壮族自治区农业科学院玉米研究所 | Method for doubling corn haploid by herbicide |
| CN109122293A (en) * | 2018-08-29 | 2019-01-04 | 广西壮族自治区农业科学院玉米研究所 | It is a kind of to double the haploid method of corn using herbicide |
| CN110192525A (en) * | 2019-06-28 | 2019-09-03 | 大连大学 | A kind of cultural method of blueberry polyploid |
| WO2021133922A1 (en) * | 2019-12-23 | 2021-07-01 | Calyxt, Inc. | Modified ploidy levels of a cannabis plant |
| CN113170727A (en) * | 2021-04-27 | 2021-07-27 | 淮阴工学院 | Breeding method of catkin willow plants |
| CN113383705A (en) * | 2021-07-19 | 2021-09-14 | 江西省农业科学院园艺研究所 | Method for breeding polyploid kiwi fruit |
| CN114041463A (en) * | 2021-11-09 | 2022-02-15 | 昆明学院 | Agents and method for reduplicating and reducing damage and improving mutagenesis rate of gynura bicolor chromosome |
| CN114521489A (en) * | 2022-03-04 | 2022-05-24 | 南通大学 | Method for inducing willow chromosome doubling at colchicine variable temperature |
| CN115669535A (en) * | 2022-10-11 | 2023-02-03 | 中国林业科学研究院经济林研究所 | Method for efficiently inducing persimmon polyploidy by treating seeds with colchicine |
| CN115500259A (en) * | 2022-10-28 | 2022-12-23 | 绍兴市农业科学研究院 | Method for overcoming pollenless of progeny of interspecific cross of pumpkin |
| CN115500259B (en) * | 2022-10-28 | 2023-08-22 | 绍兴市农业科学研究院 | Method for overcoming pollinosis of pumpkin interspecific hybridization offspring |
| CN117502225A (en) * | 2023-11-28 | 2024-02-06 | 中国热带农业科学院热带作物品种资源研究所 | Induction method and application of capsicum |
| CN117530173A (en) * | 2023-12-27 | 2024-02-09 | 中国热带农业科学院热带作物品种资源研究所 | Method for creating new germplasm of pepper tetraploid by utilizing stem tip induction |
Also Published As
| Publication number | Publication date |
|---|---|
| CN107980620B (en) | 2021-03-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN107980620A (en) | A kind of the compound of plant chromosome doubles agent and method for doubling | |
| Dhooghe et al. | Mitotic chromosome doubling of plant tissues in vitro | |
| Custers | Microspore culture in rapeseed (Brassica napus L.) | |
| Atichart | Polyploid induction by colchicine treatments and plant regeneration of Dendrobium chrysotoxum. | |
| CN103053414B (en) | Method for doubling corn haploid by herbicide and special herbicide of method | |
| CN109122293B (en) | Method for doubling corn haploid by herbicide | |
| CN102177846A (en) | Method for doubling management of corn haploid | |
| Esfahani et al. | In vitro polyploidy induction in Persian Poppy (Papaver bracteatum Lindl.) | |
| Bowman et al. | Efficient propagation of citrus rootstocks by stem cuttings | |
| CN110447534A (en) | A kind of quinoa mutation construction method based on EMS induction | |
| CN110506634A (en) | A kind of iris chemical mutagenesis dosage screening technique | |
| CN104488719B (en) | A kind of method of one-step method evoking tobacco haplobiont | |
| CN106106375A (en) | A kind of method of water planting Caulis et Folium Brassicae capitatae indoor mass rearing diamondback moth | |
| CN105104182A (en) | Method for doubling young embryos of maize haploids | |
| Iqbal et al. | Effect of pollination time on yield and quality of date fruit. | |
| Suliman et al. | Polyploidy induced by colchicine in Robinia pseudoacacia L. and it's effects on morphological, physiological and anatomical seedling traits | |
| CN106818489A (en) | A kind of lilac daphne stem section explant primary method for tissue culture | |
| CN114532225B (en) | Tissue culture rapid propagation and cultivation method for paphiopedilum delbrueckii | |
| Rhee et al. | Comparison of pollen morphology in interspecific hybrid lilies after in vitro chromosome doubling | |
| Wulandari et al. | In vitro induction of tetraploid pummelo ‘Nambangan’(Citrus maxima (Burm.) Merr.) by colchicine treatment using germinated seed, shoot tip and cotyledonary node as eplants | |
| Gülsen et al. | Growth and development of citrus pistils and fruit explants in vitro | |
| Richer et al. | Factors influencing pollen germination in three Explorer™ roses | |
| CN109874448B (en) | Method for promoting pea seed germination and seedling growth by using plant leaching liquor | |
| CN117099681B (en) | Method for doubling corn haploid by using taxol | |
| CN107410008A (en) | A kind of selection of resistance to lead Job's tears kind |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |