CN103539539A - Viola x wittrockiana chemical hybridizing agent composition and its preparation method and use - Google Patents
Viola x wittrockiana chemical hybridizing agent composition and its preparation method and use Download PDFInfo
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- CN103539539A CN103539539A CN201310476389.2A CN201310476389A CN103539539A CN 103539539 A CN103539539 A CN 103539539A CN 201310476389 A CN201310476389 A CN 201310476389A CN 103539539 A CN103539539 A CN 103539539A
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Abstract
The invention belongs to the field of flower plant leaf surface spray and relates to a viola x wittrockiana chemical hybridizing agent composition and its preparation method and use. The viola x wittrockiana chemical hybridizing agent composition comprises tribenuron-methyl, urea, gibberellin, washing powder and water. The viola x wittrockiana chemical hybridizing agent composition realizes thorough hybridization on viola x wittrockiana and normal gynoecium fertility, does not influence plant growth and development, has a chemical hybridizing rate of 96.92%, realizes normal outcrossing fructification, has an outcrossing fructification rate of 89.66% and realizes seed hybridization purity of 100%. In a spray stage, a viola x wittrockiana selfing line plant having height of 10-15cm is selected, when the largest bud size is in a range of 1-2mm, a spray process on leaf surfaces is carried out, a single-plant agent supply amount is controlled in a range of 15-20ml, and after 10 days, a spray process is carried out again and treatment concentrations at all positions are not changed and the single-plant agent supply amount is reduced to 8-10ml. The viola x wittrockiana chemical hybridizing agent composition has the advantages of good synergism, low production cost, environmental friendliness and easy popularization application.
Description
Technical field
The invention belongs to flower plant breeding technical field, be specifically related to a kind of chemical male sterilant composition and compound method and application that is applicable to wild pansy.
Background technology
Wild pansy (Viola * wittrockiana), Violaceae (Violaceae) Viola (Viola).Its flower type is lovely changeable, and pattern is gorgeous, wide in variety, and the florescence is longer, has the good reputation of " flower bed queen ", is one of preferred material of Winter-Spring flower bed, flower border, potted flower and pattern etc.; Meanwhile, because its winter hardiness is stronger, can it bloomed around New Year's Day by regulating the full-bloom stage that regulates and controls wild pansy sowing time, therefore, have a wide range of applications.
Breeding for heterosis is one of important method of cultivating at present new variety.It can make filial generation obtain parents' good character simultaneously, to reach objects such as improving the sights of flowers and resistance, for huge contribution has been made in agricultural development.But in breeding research process, often need to expend a large amount of manpower and materials, complete the emasculation work of self-pollinated plant.To with manpower, complete at the florescence emasculation of every flower, the particularly artificial emasculation of little Hua type plant, one and lengthy and tedious, time-consuming work beyond doubt.Therefore, the limited amount of cenospecies, causes breeding process to delay.This has caused great difficulty also to popularization and the application of new variety.
Male sterilant (Chemical Hybridizing Agent, CHA) gametocide is before pollen formation, interior absorption chemical agent to be sprayed on plant, suppresses pollen normal development or stops pollen formation, reaches emasculation object, for hybrid seeding provides maternal.Chemical emasculation hybrid, easy to operation, combination selection is freely, can effectively replace artificial emasculation, reduce the breeding time limit, F1 generation seed is used devoid of risk, do not need " two are " or " three are " supporting, without seed selection self incompatible line and self-mating system, be not more subject to the impact of maternal undesirable cell matter, on a large scale Selection parent yet, the shape cooperations such as high yield, high-quality are got up, greatly alleviate breeding work amount, improved breeding efficiency; Can produce at short notice the F1 generation seed of sufficient amount, be the breeding method with good prospect.
It is the important channel of plant heterosis utilization that chemical emasculation is realized assorted optimization, and the hybrid wheat that the U.S. extensively promotes at present utilizes the male sterilant production of hybrid seeds; China's research and application achievements of chemical emasculation on Cruciferae cole crop is remarkable.But the impact of chemical emasculation effect climate, plant development situation, same chemical emasculation agent concentration is for the same crop in different areas, or the gametocidal effect of areal Different Crop is different.Yet, up to now, yet there are no the special-purpose male sterilant of developing for wild pansy and the report that uses male sterilant production first generation of hybrid seed.
Based on above reason, a kind of efficient, low toxicity of research and probe, broad spectrum wild pansy male sterilant high, cheap and easy to use, making it be applied to wild pansy and utilize self-mating system production first generation of hybrid seed, is that those skilled in the art are concerned about to realize hybridizing rate 96.82%.
Summary of the invention
The object of the invention is to overcome the deficiencies in the prior art, a kind of chemical male sterilant composition and compound method and application that is applicable to wild pansy is provided.Chemical male sterilant composition of the present invention is comprised of tribenuron-methyl, urea, Plant hormones regulators,gibberellins, washing powder and water.Male sterilant prepared by the present invention is more thorough to the plant gametocidal effect of wild pansy bud phase, and gynoecium is grown normal, and plant is had no side effect.In wild pansy first generation of hybrid seed production process, adopt after male sterilant of the present invention, the florescence after testing chemical emasculation rate be 96.82%, outcrossing seed-setting rate is 89.66%.
For achieving the above object, the present invention is by the following technical solutions:
A male sterilant, is selected from following material with activeconstituents by the composition of concentration proportioning: tribenuron-methyl concentration 0.30mg/L, and urea 0.30mg/L, Plant hormones regulators,gibberellins 0.20mg/L, washing powder 0.20mg/L, keeps the skin wet to full dose (being 1L).The molecular formula of described tribenuron-methyl is C
15h
17n
5o
6s, urea molecule formula is CH
4o(analytical pure is 99.8%); Plant hormones regulators,gibberellins molecular formula is C
19h
22o
6; Washing powder main component is Sodium dodecylbenzene sulfonate (content>=13%), and molecular formula is C
18h
29naO
3s.
Applicant provides a kind of application method of composition of wild pansy male sterilant, and its step is as described below:
(1) preparation male sterilant solution:
1) preparation tribenuron-methyl solution: get 0.3g10% tribenuron-methyl, be settled to 250ml with distilled water, obtain tribenuron-methyl solution;
2) preparation urea soln: get 0.3g urea, fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains urea soln;
3) preparation Gibberellins solution: get 0.2g75% Plant hormones regulators,gibberellins (GA3), fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains Gibberellins solution;
4) preparation washing powder solution: use 0.2g washing powder, distilled water is settled to 250ml and obtains washing powder solution;
5) tribenuron-methyl solution, urea soln, Gibberellins solution, the washing powder solution of above-mentioned preparation are mixed, obtain 1L wild pansy chemical male sterilant composition.
(2) determining of spraying period:
In wild pansy squaring period, when self-mating system plant height reaches 10～15cm, during the long 1～2mm of maximum bud, utilize small manual sprayer (1L) evenly to spray wild pansy plant blade face.After 10d, once, concentration is constant in multiple spray.The time of spraying is selected in 9:00～11:00 in morning, and spraying weather should be calm, without dew, to guarantee pesticide supplying effect.
(3) individual plant is subject to dose:
Dispenser for the first time should be controlled dosage 15～20ml/ strain, sprays for the second time, and individual plant is subject to dose to reduce to 8～10ml/ strain.
Beneficial effect of the present invention is:
The chemical emasculation composition advantage of using in the present invention is, plant is had no side effect, and can induce wild pansy to produce plant male sterile thoroughly, and gynoecium is grown normal; Utilize male sterilant to reach 96.82% to the hybridizing rate of wild pansy.The present invention is environmentally friendly, and wherein tribenuron-methyl is a kind of conventional chemical herbicide, extensively relates to for agriculture production; Urea is a kind of neutral fertilizer, on soil, without impact, is a kind of high-quality and efficient nitrogenous fertilizer; Plant hormones regulators,gibberellins is the class plant hormone extensively existing, and can stimulate the growth of leaf and bud; Washing powder is a kind of conventional surfactant, good and cheap; Above-mentioned chemical substance is non-heavy metal compounds, and toxicity is low, residual few, to person poultry safety; In soil, easily degraded, safer to environment; Active high, consumption is few, and cost is low, easy to use, benefits popularization.
Accompanying drawing explanation
Fig. 1: be to not spraying the observation of No. 21 normal Pollen Activities of wild pansy angle violet of chemical male sterilant composition with 1% acetic red dyeing.
Fig. 2: be with 1% acetic red dyeing, wild pansy angle violet to be sprayed the observation of the Pollen Activity after chemical male sterilant composition for No. 21.
Fig. 3: be the normal pollen germination situation that wild pansy angle violet does not spray chemical male sterilant composition for No. 21.
Fig. 4: be that wild pansy angle violet sprays the pollen germination situation after chemical male sterilant composition for No. 21.
Fig. 5: be the upgrowth situation that wild pansy angle violet does not spray the normal plant of chemical male sterilant composition for No. 21.
Fig. 6: be the upgrowth situation that wild pansy angle violet sprays plant after chemical male sterilant composition for No. 21.
Fig. 7: be the morphologic observation that wild pansy angle violet does not spray the normal flower of chemical male sterilant composition for No. 21.
Fig. 8: be the morphologic observation that wild pansy angle violet sprays flower after chemical male sterilant composition for No. 21.
Fig. 9: be the wild pansy angle violet pollen tube growth of normal flower after outcrossing pollination that do not spray chemical male sterilant composition for No. 21 and the situation that enters ovary.
Figure 10: be that wild pansy angle violet sprays pollen tube growth after outcrossing pollination of flower after chemical male sterilant composition for No. 21 and enters the situation of ovary.
Embodiment
Be below the embodiment that contriver provides, it should be noted that, example, only for explaining the present invention, is not intended to limit protection scope of the present invention.
Embodiment 1:
The single medicament of tribenuron-methyl sprays, and concentration is respectively 0.00,0.30,0.50,0.70ug/mL.At in mid or late March, 2012, No. 21 self-mating system plant height reaches 10～15cm, during the long 1～2mm of maximum bud, utilizes small manual sprayer (1L) evenly to spray wild pansy plant blade face.Individual plant is subject to dose to be controlled at 15～20mL, sprays again 1 time after 10d, and each concentration for the treatment of is constant, and dose is reduced to 8～10mL.10 strains are a processing, process 1 time, spray clear water and compare.At the florescence, observe gametocidal effect and the tribenuron-methyl of wild pansy and process the impact on pansy flowers organ.
The gametocidal effect of table 1 different concns tribenuron-methyl to wild pansy
0.30,0.50 and 0.70 μ g/mL tribenuron-methyl is processed 1 energy induction of male sterility flower ratio and is respectively 86.95%, 96.90%, 100%.0.30,0.50 and 0.70 μ g/mL processes from flower self-fruitful rate and is respectively 6.67%, 3.33%, 0.00%, is starkly lower than 96.67% of fertile plant (0.00 μ g/mL).And the processing outcrossing seed-setting rate of 0.3 μ g/mL is 93.33%, compares and there is no notable difference with fertile plant (0.00 μ g/mL).The processing outcrossing seed-setting rate of 0.5 and 0.7 μ g/mL is respectively 73.33% and 0.00%.This explanation, tribenuron-methyl more than 0.50 μ g/mL is processed and has not only been induced pollen abortion, also gynoecium has been produced to larger injury (illustrating: in table 1, the gametocidal effect of wild pansy calculates according to pollen germination rate).
The impact of table 2 different concns tribenuron-methyl on pansy flowers organ morphology
Compare with fertile plant (0.00 μ g/mL), tribenuron-methyl is spent all obviously cripeturas of footpath, gynoecium, column cap, flower pesticide after processing.But 0.3 μ g/mL concentration is processed and fertile plant (0.00 μ g/mL) no significant difference.After tribenuron-methyl is processed, corolla reduces, and epivalve pattern fades to lilac by intense violet color, hypovalve major part becomes white, only has part fickle in love for yellow, and mesopetalum circle fades to rectangle, and concentration is larger, change more obviously, 0.70 μ g/mL concentration is processed whole petal and is all lavender; Flower pesticide is shrivelled, and shortening broadens, and Pollen Activity is little.0.50,0.70 μ g/mL concentration is processed column cap anamorphosis, and ovary obviously reduces.
Embodiment 2:
1, material
Test materials adopts wild pansy new variety ' ice is purple ' (being not limited to the kind of the present embodiment in concrete embodiment), these new variety are published, referring to network address: http://www.pyyl.net/zhuantibaodao/2011-11-10/18792.html; Four > > of the 29 national woods scientific and technical information network net of < < meeting, gardens, Puyang net is reprinted, deliver these new variety of time 2011-11-10(authorizes by the Hubei Province forest variety certification council for 2010, to be formed with No. 22 (for conventional variety) hybridization of great Hua wild pansy for No. 21 by angle violet), this kind plant type is compact, plant height 20～30cm, branch amount 18～25, after autumn sowing, 90～95d is bloomed.
The present invention sprays with hand formula atomizer; With 10cm * 5cm sulfuric acid paper bag to processing the spray bagging of strain and according to the ordinary method isolation production of hybrid seeds.
2, test method
(1) field planting of growing seedlings of the Parent of production first generation of hybrid seed
By the Parent of wild pansy seeding and seedling raising at the beginning of 10 months then, sowing medium answers draining good, loose.After broadcasting, need tightly to cover 1cm left and right thin soil, and be placed in lucifuge place.After broadcasting, to water permeablely, note moisturizing.Germination thermophilic is 15～20 ℃.When wild pansy starts to grow after first pair of true leaf, give sufficient sunlight, in order to avoid excessive growth.When growing to 2 pairs of true leaves, seedling to transplant immediately, carry out potted plant or outdoor cropping, generally adopt basin on the nutrition pot of 12 * 13cm, for garden mould+leaf-humidified soil+yellow sand, by weight 3:2:1 preparation, (described garden mould is common vegetable garden soil to basin soil, do not add artificial feeding material), Routine Management.
(2) tribenuron-methyl chemical male sterilant composition is processed, and orthogonal experimental design is as follows:
Table 3 orthogonal experiment L
9(3
4) level of factor table
Take 2 levels as example, and chemical male sterilant composition compound method is as follows:
1) preparation tribenuron-methyl solution: get 0.3g10% tribenuron-methyl, be settled to 250ml with distilled water, obtain tribenuron-methyl solution.
2) preparation urea soln: get 0.3g urea, fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains urea soln.
3) preparation Gibberellins solution: get 0.2g75% Plant hormones regulators,gibberellins (GA3), fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains Gibberellins solution.
4) preparation washing powder solution: use 0.2g washing powder, distilled water is settled to 250ml, obtains washing powder solution.
5) tribenuron-methyl solution, urea soln, Gibberellins solution and the washing powder solution of above-mentioned preparation are mixed, obtain 1L wild pansy chemical male sterilant composition.
(3) maternal plant sprays male sterilant period, dose and method
In next year, at maternal angle, violet plant height reaches 10～15cm, during the long 1～2mm of maximum bud, utilizes violet plant blade face, small manual sprayer diagonal angle evenly to spray.After 10d, once, concentration is constant in multiple spray.Using 10 strains as a processing, to spray distilled water in contrast.Spray time 9:00～11:00 in the morning, spray weather calm without dew, guarantee drug effect; Individual plant is sprayed for the first time by dose and is controlled at 15～20ml, sprays for the second time, and individual plant is subject to dose to reduce to 8～10ml.
(4) production of first generation of hybrid seed
After Parent flowering, male parent floral organ is normal, on No. 21 flower column caps of maternal angle violet that the pollen that No. 22, collection male parent is pollinated after gametocide, and production first-filial generation seed on the maternal plant after gametocide.
(5) first generation of hybrid seed testing
After the violet seed maturity of maternal angle, the seed of gathering in maternal strain carries out field hybridization Purity.
3, interpretation of result
Table 4 tribenuron-methyl combination L
9(3
4) orthogonal scheme and interpretation of result
In four concentration, tribenuron-methyl concentration and Plant hormones regulators,gibberellins concentration affects are comparatively remarkable, and wherein tribenuron-methyl concentration affects is the most remarkable.Optimum chemical male sterilant composition is: tribenuron-methyl concentration 0.30 μ g/mL, urea 0.30 μ g/mL, Plant hormones regulators,gibberellins 0.20 μ g/mL, washing powder 0.20 μ g/mL.
Under this optimal set compound is processed:
1) change and kill hybridizing rate, outcrossing seed-setting rate and hybrid seed purity
Chemical emasculation rate is 96.82%(explanation: the gametocidal effect of the wild pansy of the present embodiment calculates according to pollen germination rate), outcrossing seed-setting rate is 89.66%, hybrid seed purity is 100%.
2) impact of chemical emasculation on plant forms and floral organ
On plant forms, terminal bud leaf look shoals, and plant height slightly reduces; On flower organ morphology, corolla reduces, and pattern shoals, and pollen sky is flat, flower pesticide anamorphosis.Gynoecium is uninfluenced.
Embodiment 3:
1, material
Test materials is selected wild pansy new variety ' cat ', (being not limited to the kind of the present embodiment in concrete embodiment) these new variety are published, referring to network address: http://www.pyyl.net/zhuantibaodao/2011-11-10/18792.html; Four > > of the 29 national gardens scientific and technical information network net meeting of < <, gardens, Puyang net is reprinted; Delivering these new variety of time 2011-11-10(' cat ' and by the Hubei Province forest variety certification council, authorize for 2010, is by No. 21, angle violet and No. 9 (for conventional variety) cross breedings of wild pansy).With 10cm * 5cm sulfuric acid paper bag to processing the spray bagging of strain and according to the ordinary method isolation production of hybrid seeds.
The present invention sprays with hand formula atomizer.
2, test method
(1) field planting of growing seedlings of the Parent of production first generation of hybrid seed
Parent is seeding and seedling raising at the beginning of 10 months then, and sowing medium answers draining good, loose; After broadcasting, need tightly to cover 1cm left and right thin soil, and be placed in lucifuge place; After broadcasting, to water permeablely, note moisturizing; Germination thermophilic is 15～20 ℃.Start to grow after first pair of true leaf, give sufficient sunlight, in order to avoid excessive growth.When growing to 2 pairs of true leaves, seedling to transplant immediately, carry out potted plant or outdoor cropping, generally adopt basin on the nutrition pot of 12 * 13cm, for garden mould+leaf-humidified soil+yellow sand, by weight 3:2:1 preparation, (described garden mould is common vegetable garden soil to basin soil, do not add artificial feeding material), Routine Management.
(2) chemical male sterilant composition of wild pansy is prepared according to the following steps:
1) preparation tribenuron-methyl solution: get 0.3g10% tribenuron-methyl, be settled to 250ml with distilled water, obtain tribenuron-methyl solution;
2) preparation urea soln: get 0.3g urea, fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains urea soln;
3) preparation Gibberellins solution: get 0.2g75% Plant hormones regulators,gibberellins (GA3), fully dissolve with 75% ethanol, distilled water is settled to 250ml, obtains Gibberellins solution;
4) preparation washing powder solution: use 0.2g washing powder, distilled water is settled to 250ml, obtains washing powder solution;
5) tribenuron-methyl solution, urea soln, Gibberellins solution, the washing powder solution of above-mentioned preparation are mixed, obtain 1L wild pansy chemical male sterilant composition.
(3) maternal plant sprays male sterilant period, dose and method
In next year, at maternal angle, violet plant height reaches 10～15cm, during the long 1～2mm of maximum bud, utilizes violet plant blade face, small manual sprayer (1L) diagonal angle evenly to spray.After 10d, once, concentration is constant in multiple spray.Using 10 strains as a processing, to spray distilled water in contrast.Spray time 9:00～11:00 in the morning, spray weather calm without dew, guarantee drug effect; Individual plant is sprayed for the first time by dose and is controlled at 15～20mL, sprays for the second time, and individual plant is subject to dose to reduce to 8～10ml.
(4) production of first generation of hybrid seed
After Parent flowering, male parent floral organ is normal, on No. 9 flower column caps of maternal wild pansy that the pollen that No. 21, collection male parent is pollinated after gametocide, and production first-filial generation seed on the maternal plant after gametocide.
(5) first generation of hybrid seed testing
After maternal wild pansy ninth-seeded maturation, the seed gathering in maternal strain carries out field hybridization Purity.
The chemical emasculation rate of the present embodiment is 96.82%, and outcrossing seed-setting rate is 89.66%, and hybrid seed purity is 100%.
(6) impact of chemical emasculation on plant forms and floral organ
On plant forms, terminal bud leaf look shoals, and plant height slightly reduces; On flower organ morphology, pollen sky is flat, flower pesticide anamorphosis.Gynoecium is uninfluenced.
Utilize the technology of the present invention to carry out " emasculation is killed in change " to the fertile plant of wild pansy, in the situation that female parent is identical, greatly saved the production of hybrid seeds time, simplified production of hybrid seeds step, F1 generation purity and advantage no significant difference, production of hybrid seeds expense declines to a great extent; " it " is a kind of innovation of wild pansy emasculating method that emasculation is killed in change.
Claims (3)
1. a chemical male sterilant composition that is applicable to wild pansy, is characterized in that, as described below by the component of mg/L: tribenuron-methyl concentration 0.30mg/L, and urea 0.30mg/L, Plant hormones regulators,gibberellins 0.20mg/L, washing powder 0.20mg/L, surplus is water.
2. a compound method that is applicable to the chemical male sterilant composition of wild pansy, is characterized in that the following step:
1) preparation tribenuron-methyl solution: get 0.3g10% tribenuron-methyl, be settled to 250ml with distilled water, obtain tribenuron-methyl solution;
2) preparation urea soln: get 0.3g urea, first fully dissolve with 75% ethanol, then be settled to 250ml with distilled water, obtain urea soln;
3) preparation Gibberellins solution: get 0.2g75% Plant hormones regulators,gibberellins GA3, first fully dissolve with 75% ethanol, then be settled to 250ml with distilled water and obtain Gibberellins solution;
4) preparation washing powder solution: first use 0.2g washing powder, then be settled to 250ml with distilled water and obtain washing powder solution.
5) tribenuron-methyl solution, urea soln, Gibberellins solution, the washing powder solution of above-mentioned preparation are mixed, obtain 1L wild pansy chemical male sterilant composition.
3. apply a method for wild pansy chemical male sterilant composition as claimed in claim 1, it is characterized in that the following step:
In wild pansy squaring period, when self-mating system plant height is 10～15cm, the long 1～2mm of maximum bud, evenly sprays wild pansy plant blade face, and formulation rate is 15～20ml/ strain, and after 10d, once, formulation rate is 15～20ml/ strain in multiple spray.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1524417A (en) * | 2003-02-28 | 2004-09-01 | 陕西省杂交油菜研究中心 | Chemical crossbred agent composition, its using method and application |
| CN101473838A (en) * | 2009-02-11 | 2009-07-08 | 西北农林科技大学 | Chemical anthericide for cabbage stalk and formulating method thereof |
| CN102440136A (en) * | 2011-11-18 | 2012-05-09 | 大连好地农业有限公司 | Hybrid pansy seed production method |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1524417A (en) * | 2003-02-28 | 2004-09-01 | 陕西省杂交油菜研究中心 | Chemical crossbred agent composition, its using method and application |
| CN101473838A (en) * | 2009-02-11 | 2009-07-08 | 西北农林科技大学 | Chemical anthericide for cabbage stalk and formulating method thereof |
| CN102440136A (en) * | 2011-11-18 | 2012-05-09 | 大连好地农业有限公司 | Hybrid pansy seed production method |
Non-Patent Citations (1)
| Title |
|---|
| 于爱霞: "三色堇自交系遗传多样性评价及化学去雄的初步研究", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
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