CN104604673A - Treatment method for inducing maize parthenogenesis with colchicine - Google Patents
Treatment method for inducing maize parthenogenesis with colchicine Download PDFInfo
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- CN104604673A CN104604673A CN201410405142.6A CN201410405142A CN104604673A CN 104604673 A CN104604673 A CN 104604673A CN 201410405142 A CN201410405142 A CN 201410405142A CN 104604673 A CN104604673 A CN 104604673A
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Abstract
The invention relates to a treatment method for inducing maize parthenogenesis with colchicine. The method is characterized in that when female ear filaments are completely exposed, a paper bag is removed; the upper-portion of a flower bud is cut transversely from a female ear flower bud at a place from the top and close to the female ear young ear downward; a part of filaments is cut at the section with a curved graver, such that a cavity with a depth of 5-15mm is formed; an inducing agent which is colchicine treatment liquid is injected into the cavity with a pipette or a syringe, such that the cavity is filled with the liquid and the entire cut surface is wetted by the liquid; and the female ear is sleeved with the paper bag. During the treatment process, no pollen contamination prevention measure such as sealing, blocking, spatial and temporal isolation, and the like is needed. The method has the following advantages: harm to human health caused by treatment liquid dropping, spilling and scratching are avoided; a hard-to-solve pollen contamination problem is thoroughly solved; all filaments can contact the treatment liquid, and treatment liquid retention time is long, such that an induction rate is improved; the treatment liquid is saved by approximately 50%; and compared to a conventional inbred line breeding method, a breeding period is shortened by more than 5 generations.
Description
Technical field
The invention belongs to corn breeding techniques field, particularly a kind of selection of corn inbred line.
Background technology
The conventional method of selecting and breeding corn inbred line, be bred as the continuous selfing that an inbred line at least needs more than 7 generations, 10 generations more than even wanted by the material for genetic background complexity.In order to accelerate the seed selection speed of inbred line, shorten breeding cycle and improve purity, the method usually adopted both at home and abroad mainly contains:
1. colchicine-induced method
In the corn silking phase, with colchicine as chromosome doubling inorganic agent process filigree, make it without double fertilization but directly produced the dliploid seed isozygotied by parthenogenesis.
Colchicine is conventional chromosome doubling inorganic agent, and its outstanding advantages is that inductivity is high, is therefore widely used in teaching and scientific research.But, although the parthenogenesis of colchicine-induced corn has been a kind of corn inbred lines technology of maturation.But the two large problems that the method exists, makes it be difficult to scale application in breeding:
One is safety problem.Colchicine belongs to poisonous reagent, and carcinogenicity is strong.In order to avoid and human contact as far as possible, prior art is all taked to smear filigree or the method to filigree injection.But very easily cause scratching and liquid drippage when smearing, because the space between filigree is very little during injection, consumption is difficult to grasp, and very easily causes liquid medicine spill and trickling.Therefore, threaten very large to health.
Two is pollen contamination problems.During process, even if operate under strict bagging condition, or block, or avoid the loose powder time, but pollen contamination problem is still very serious, produces a large amount of pseudosperm, particularly under the weather conditions of Hainan.
In addition, because filigree growth is tight, little to the saturation of liquid during injection, a part of filigree is not easy to touch liquid; Again because filigree hydrophily is poor, be not easy attachment when smearing, be difficult to reach desirable treatment effect; In order to ensure safety, operation requirements is extremely strict.Therefore, process action careful, speed is slow, efficiency is low, be also cause can not scale application reason.
2. revulsion
The pollination of derivative object is given, inducing maize Haploid production seed with genetic induction system Stock 6.Liploid plant is doubled into more afterwards with chromosome doubling inorganic agents such as colchicines.The method is considered at present state-of-the-art method in the world.But, still there is following problem:
(1) the monoploid seed of induction generation, needs to utilize mark property to screen by grain the seed on whole tassel, selects the monoploid seed produced by induction, eliminates double fertilization seed.Therefore, workload is large, efficiency is low, hand picking is dry as dust, is difficult to realize mechanization;
(2) for part induction object, due to reasons such as genetic background, maturity, Seed shape, peel thickness, seed adnation positions, also there is the not good problem even do not developed the color of color developing effect, be therefore difficult to identification;
(3) induce the monoploid seed produced just must can become normal dliploid through chromosome doubling again with Stock 6, and the process that doubles is loaded down with trivial details and to add multiplying power lower;
(4) when utilizing colchicine to carry out chromosome doubling, the waste liquid after process belongs to severe toxicity, carcinogen, needs strict harmless treatment;
(5) use Stock 6 genetic induction method, need artificial pollination.In order to make induction system and the flower synchronization being induced object, intermission seeding be carried out, regulating father, maternal flowering time.But, often because the reaction of test material to light, temperature is different, or because of reasons such as soil, fertility differences, easily there is flowering asynchronism.
3. other reagent revulsion
Process filigree with such as the trefanocide etc. of other chemical reagent with induce chromosome redoublement effect of low toxicity, induced parthenogenesis directly produces dliploid seed.Compared with Stock 6 revulsion, have person poultry safety, method is simple, processing ease, cost are low, efficiency advantages of higher.
But compared with colchicine, the inductivity of these reagent is relatively low, therefore need to strengthen process quantity.Workload is comparatively large, and cost improves.
Summary of the invention
The technical problem to be solved in the present invention is: (1) safe handling colchicine inorganic agent.The advantage of colchicine height inductivity should be utilized, in method of operating, solve the safety problem that existing method exists again; (2) problem that in the existing method of chemical induction, ubiquitous pollen contamination rate is high, pseudosperm is many is solved; (3) simplify processes program, make simple to operate, be easy to scale application; (4) improve inductivity simultaneously, reduce dosing.
The technical solution used in the present invention is as follows:
1. derivant preparation
Colchicine (COL) derivant that preparation is conventional, or with colchicine be main with dimethyl sulfoxide (DMSO) (DMSO), the composite derivant of maleic hydrazide (MH).
2. fruit ear process before induction
To female fringe bagging before weaving silk; When filigree spues completely, remove paper bag, divide petal close to the horizontal cut away in position of female fringe children fringe at female Honoka bud downwards by top.Presenting centre is filigree, is around the circular cross section of bract; On section, cut away one section of filigree with arc icking tool, the degree of depth of cutting is 8-15mm, accomplishes not only to cut away filigree but also do not cut the bract Lou, and namely on section, Formation Depth is the cavity of 8-15mm.
3. inject derivant
For preventing medicament and human contact, derivant is loaded tight-closed utensil as liquid-transfering gun or syringe.Backward cavity in inject derivant, to be full of and liquid covers or till wetting whole section.Finally paper bag is recovered, complete processing procedure.
Due to pollen runs into after treatment fluid can the death of expanding because absorbing liquid, even if section has pollen fall into also can not cause any pollution.Therefore, thoroughly solve pollen contamination problem, guarantee that seed is true and reliable.During process, do not need sealing, block or isolate, whole process is free operant under complete open condition.
4. results and breeding
Gather in the crops during seed maturity, namely obtain the dliploid seed that genotype is isozygotied; Next season is sowed, bagging selfing, carries out the true and false and good and bad qualification, select elite plant, obtain single fringe according to phenotype.And it can make male parent and be measured in advance in this generation; The fruit ear of results is continued sowing of future generation, plant into head progeny row, the uniformity according to proterties such as plant, fruit ear, seeds identifies the head progeny row that isozygotys, and selects the head progeny row results meeting breeding objective, namely obtains the corn inbred line isozygotied.Can be measured in a large number in this generation.
Good effect of the present invention is as follows:
1. solve the safety problem using colchicine
Cross section is adopted to dig a hole dispenser, the scratching caused when avoiding prior art owing to smearing and liquid drippage, the threat that liquid medicine spill during injection, trickling etc. cause health.
2. solve the pollen contamination problem of prior art
Flood the top of whole filigree due to treatment fluid and covered whole tangent plane, also can expand dead because pollen runs into water suction after treatment fluid even if there is pollen to descend slowly and lightly, thoroughly solve insoluble pollen contamination problem in prior art.
3. can under complete open condition free operant
Do not need at bagging, block during induction process, space isolated area or time isolation condition, can under complete open condition and at any time in free operant.
4. be applicable to scale application
Because use safety, program are easy, easily simple to operate, and do not limit by isolated area, therefore can scale application.
5. inductivity improves
Both make use of the advantage that colchicine-induced rate is high, and ensured that again all filigrees can contact with liquid, also make liquid extended residence time, thus inductivity improves.
6. save liquid
Liquid is concentrated, and waste of not trickling.The general every fringe of existing colchicine treatment technology needs liquid about 2ml, and the method only needs about 1ml, can save liquid about 50%, reduces drug cost.
7. program is simple
Directly form dliploid seed.Compared with inducing monoploid technology with Stock 6, one is do not need chromosome doubling process; Two is do not need contemporary seeds idenmtification; Three is there is not the problems such as seeds idenmtification difficulty.
8. seed selection speed is fast
More than seed selection cycle time 5 generation of ordinary maize inbred line breeding method.Owing to can guarantee to induce the authenticity of setting seeds and induction first generation plant can normal loose powder, therefore just can it be measured in advance as male parent from the induction first generation, breeding hybridized speed induces monoploid technology also will shift to an earlier date for 1 generation than Stock 6.
Embodiment
Embodiment 1
(1) derivant is prepared
Colchicine (COL) is diluted with water to the derivant of 0.1% concentration.
(2) process examination material is selected
Handling object is corn hybrid seed " the Liao Dynasty single 565 ".
(3) fruit ear process before induction
To female fringe bagging before weaving silk.Paper bag is taken after filigree spues completely, laterally cut away top petal close to the position of female fringe children fringe at female Honoka bud downwards by top, on section, cut away one section of filigree with arc icking tool, the degree of depth of cutting is 5-15mm, do not cut the bract Lou, Formation Depth is the cavity of 5-15mm.
(4) derivant is applied
Inject derivant with liquid-transfering gun to cavity, be full of and cover or wetting whole tangent plane.Finally recover paper bag.Process 60 fringes.
Operating condition: operating process is carried out under complete open condition, does not namely need to seal, blocks, the measure preventing pollen contamination of time or space isolation.
(5) gather in the crops
Time ripe, results fruit ear, obtains 90 seeds, carries out mark.
(6) qualification and breeding
In next season by seed monoseeding, eliminate the individual plant not meeting breeding goal, bagging selfing, obtains 90 fruit ears.Be measured as male parent with it simultaneously; The fruit ear of results is continued sowing of future generation, plant into 60 head progeny rows, the strain of every head progeny row 50, bagging selfing, is measured simultaneously.According to the regularity of the proterties such as plant, fruit ear, seed, whether proterties meets breeding goal, and is measured the performance of combination and selects excellent.To select and remain 50 head progeny rows, i.e. 50 parts of inbreds isozygotied.So far the Breeding Process of inbred line is completed.
Embodiment 2
(1) derivant is prepared
Select colchicine and dimethyl sulfoxide (DMSO) (DMSO) that market is sold, carry out composite derivant by following part by weight: colchicine 0.2%, dimethyl sulfoxide (DMSO) 2%, surplus is water.
(2) process examination material is selected
Handling object is sweet corn hybrid " No. 6, Shen Tian ".
(3) fruit ear process before induction
Before weaving silk, strict bagging is carried out to female fringe.Paper bag is taken when filigree spues completely, laterally cut away top petal close to the position of female fringe children fringe at female Honoka bud downwards by top, then on section, cut away one section of filigree with arc icking tool, the degree of depth of cutting is 5-15mm, do not cut the bract Lou, Formation Depth is the cavity of 5-15mm.
(4) derivant is applied
Inject derivant with liquid-transfering gun to cavity, be full of and cover or wetting whole tangent plane.Finally recover paper bag.Process 50 fringes.
Operating condition: operating process is carried out under complete open condition, does not namely need to seal, blocks, the measure preventing pollen contamination of time or space isolation.
(5) gather in the crops
Time ripe, results fruit ear, carefully peels bract off, takes off seed, obtain 70 seeds, carry out mark.
(6) qualification and breeding
In next season by seed monoseeding, eliminate the individual plant not meeting breeding goal, bagging selfing, obtains 70 fruit ears.Be measured as male parent with it simultaneously; The fruit ear of results is continued sowing of future generation, plant into 70 head progeny rows, the strain of every head progeny row 40, bagging selfing.Be measured simultaneously.According to the regularity of the proterties such as plant, fruit ear, seed, whether proterties meets breeding goal, and is measured the performance of combination and selects excellent.To select and remain 50 head progeny rows, i.e. 50 parts of inbreds isozygotied.So far the Breeding Process of inbred line is completed.
Embodiment 3
(1) derivant is prepared
Select colchicine, dimethyl sulfoxide (DMSO) and maleic hydrazide (MH) that market is sold, carry out composite derivant by following part by weight: colchicine 0.2%, dimethyl sulfoxide (DMSO) 2%, 40mgkg-1 maleic hydrazide, surplus is water.
(2) process examination material is selected
Handling object is corn hybrid seed " first beautiful 335 ".
(3) fruit ear process before induction
To the strict bagging of female fringe before weaving silk.Paper bag is taken when filigree spues completely, laterally top petal is cut away close to the position of female fringe children fringe downwards by top at female Honoka bud, form circular cross-section, on section, one section of filigree is dug up again with arc icking tool, the degree of depth of cutting is 5-15mm, do not cut the bract Lou, Formation Depth is the cavity of 5-15.
(4) derivant is applied
Inject derivant with liquid-transfering gun to cavity, be full of and cover or wetting whole tangent plane.Finally recover paper bag.Process 50 fringes.
Operating condition: operating process is carried out under complete open condition, does not namely need to seal, blocks, the measure preventing pollen contamination of time or space isolation.
(5) gather in the crops
Gather in the crops fruit ear after maturation, gather in the crops 70 seeds, mark.
(6) qualification and breeding
Next season, eliminate the individual plant not meeting breeding goal, bagging selfing, obtained 55 fruit ears by seed monoseeding.Be measured as male parent with it simultaneously; The fruit ear of results is continued sowing of future generation, plant into 55 head progeny rows, the strain of every head progeny row 50, bagging selfing.Be measured simultaneously.According to the regularity of the proterties such as plant, fruit ear, seed, whether proterties meets breeding goal, and is measured the performance of combination and selects excellent.To select and remain 40 head progeny rows, i.e. 40 parts of inbreds isozygotied.So far the Breeding Process of inbred line is completed.
Embodiment 4
(1) derivant is prepared
Colchicine that market is sold and dimethyl sulfoxide (DMSO) is selected to carry out composite derivant: the derivant being re-dubbed colchicine concentration 0.1%+ dimethyl sulfoxide concentration 0.1%.
(2) process examination material is selected
Handling object is pop corn crossbreed " quick-fried No. 3 of Shen ".
(3) induction process
To the strict bagging of female fringe before weaving silk.Paper bag is taken after filigree spues completely, laterally top petal is cut away close to the position of female fringe children fringe downwards by top at female Honoka bud, form a circular cross-section, on section, upper part filigree is dug up again with arc icking tool, the degree of depth of cutting is 5-15mm, do not cut the bract Lou, Formation Depth is the cavity of 5-15mm.
(4) derivant is applied
Inject derivant with liquid-transfering gun to cavity, be full of and soak whole tangent plane.Finally recover paper bag.Process 60 fringes.
Operating condition: operating process is carried out under complete open condition, does not namely need to seal, blocks, the measure preventing pollen contamination of time or space isolation.
(5) gather in the crops
Gather in the crops fruit ear after maturation, obtain 80 seeds, carry out mark.
(6) qualification and breeding
In next season by seed monoseeding, eliminate the individual plant not meeting breeding goal, bagging selfing, obtains 45 fruit ears.Be measured as male parent with it simultaneously; The fruit ear of results is continued sowing of future generation, plant into 45 head progeny rows, the strain of every head progeny row 50, bagging selfing.Be measured simultaneously.According to the regularity of the proterties such as plant, fruit ear, seed, whether proterties meets breeding goal, and is measured the performance of combination and selects excellent, 35 head progeny rows of preferentially selecting and remain, i.e. 35 parts of inbreds isozygotied.So far the Breeding Process of inbred line is completed.
Process examination material used in the specific embodiment of the invention is the corn seed of market sale.
Claims (1)
1., by the gynecogenic processing method of colchicine-induced corn, it is characterized in that:
(1) pre-treatment fruit ear is induced: when female Honoka silk spues completely, take paper bag, laterally the upper part of petal is cut away close to the position of female fringe children fringe downwards by top at female Honoka bud, filigree in the middle of being formed, be around the circular cross section of bract, then on section, dig up one section of filigree with arc icking tool, depth of cut is 5-15mm, do not cut the bract Lou, Formation Depth is the cavity of 5-15mm;
(2) derivant is injected: inject colchicine inorganic agent liquid with liquid-transfering gun or syringe to cavity, be full of, and make liquid soak tangent plane, finally recover paper bag;
(3) operating condition: operating process is carried out under complete open condition, does not namely need to seal, blocks, the measure preventing pollen contamination of time or space isolation.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410405142.6A CN104604673B (en) | 2014-08-18 | The gynecogenic processing method of one colchicine-induced Semen Maydis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410405142.6A CN104604673B (en) | 2014-08-18 | The gynecogenic processing method of one colchicine-induced Semen Maydis |
Publications (2)
| Publication Number | Publication Date |
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| CN104604673A true CN104604673A (en) | 2015-05-13 |
| CN104604673B CN104604673B (en) | 2017-01-04 |
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Cited By (5)
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| CN105918115A (en) * | 2016-05-03 | 2016-09-07 | 石河子农业科学研究院 | Method for obtaining pure line through inducing parthenogenesis of Zea mays by chemical agent |
| CN105941140A (en) * | 2016-05-22 | 2016-09-21 | 沈阳农业大学 | Chemical induction corn parthenogenesis operation method |
| CN106577263A (en) * | 2016-12-19 | 2017-04-26 | 陕西省杂交油菜研究中心 | Precision application method for chemical hybridization agent for rapes |
| CN112772411A (en) * | 2021-02-24 | 2021-05-11 | 内蒙古民族大学 | Breeding method for inducing maize apomixis by using mixed reagent and application |
| CN115812594A (en) * | 2022-12-29 | 2023-03-21 | 湖南农业大学 | Efficient breeding method for inducing parthenogenesis of corn by using chemical reagent |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105918115A (en) * | 2016-05-03 | 2016-09-07 | 石河子农业科学研究院 | Method for obtaining pure line through inducing parthenogenesis of Zea mays by chemical agent |
| CN105941140A (en) * | 2016-05-22 | 2016-09-21 | 沈阳农业大学 | Chemical induction corn parthenogenesis operation method |
| CN106577263A (en) * | 2016-12-19 | 2017-04-26 | 陕西省杂交油菜研究中心 | Precision application method for chemical hybridization agent for rapes |
| CN106577263B (en) * | 2016-12-19 | 2019-04-05 | 陕西省杂交油菜研究中心 | A kind of rape chemical hybridizing agent accuracy pesticide applying method |
| CN112772411A (en) * | 2021-02-24 | 2021-05-11 | 内蒙古民族大学 | Breeding method for inducing maize apomixis by using mixed reagent and application |
| CN112772411B (en) * | 2021-02-24 | 2023-03-24 | 内蒙古民族大学 | Breeding method for inducing maize apomixis by using mixed reagent and application |
| CN115812594A (en) * | 2022-12-29 | 2023-03-21 | 湖南农业大学 | Efficient breeding method for inducing parthenogenesis of corn by using chemical reagent |
| CN115812594B (en) * | 2022-12-29 | 2023-11-24 | 湖南农业大学 | Efficient breeding method for inducing parthenogenesis of corn by using chemical reagent |
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