CN1994065A - Method for improving seed selection of watermelon polyploid - Google Patents
Method for improving seed selection of watermelon polyploid Download PDFInfo
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Abstract
The invention relates to a method for improving the polyploidy cultivation of water melon, wherein it comprises that: immerging diploid seed into normal-temperature water for 6-8h; immerging it with 0.05-0.1% colchicines for 16-20h; using wet towel to pack and putting it into cultivate box at 28-32Deg. C to bud for 48-72h; planting into cavity disc; cultivating in green room; putting seed into artificial intelligent cultivate box at 12-18De. g C, taking off the growing point; processing with colchicines, dropping it one time each day, processing for 4-6 days; inducing polyploidy, growing new growing point, checking polyploidy, pollinating, and storing the seed. The invention can improve induce rate of polyploidy water melon.
Description
Technical field
The invention belongs to the agricultural crops breeding technical field, relate to a kind of method that improves seed selection of watermelon polyploid.
Background technology
For obtaining good TRIPLOID SEEDLESS WATER kind, obtaining the tetraploid watermelon material by chromosome doubling is the key of breeding success
[1,2]The China and the U.S. have now become the major country of research and cultivation TRIPLOID SEEDLESS WATER
[2,3]Produce aspermous watermelon with polyploid and become main path.TRIPLOID SEEDLESS WATER is to utilize one of plant variety of autopolyploid area maximum at present on producing
[4]
The formation approach of watermelon polyploid has artificial induction and two kinds of main modes of abiogenesis.The artificial induction doubles the zygotic number of original seed or hybrid by the whole bag of tricks.And abiogenous frequency is very low, is that fertilization by original seed or unreduced gamete that hybrid forms is in conjunction with chromosome number is doubled.The artificial induction obtains the tetraploid watermelon watermelon following approach: (1) utilizes physical method to induce: utilize induce variations such as various rays, abnormal temperature, ultracentrifugation power, high voltage.This kind method variation direction is more, but the induced mutation rate probability is very low.(2) biological method: as tetraploid occurring at the callus place after repeatedly pinching, the grafting.The induced mutation rate of this kind method is also very low, does not generally adopt.(3) utilize chemical method to induce: to utilize chemical reagent such as colchicine, vinylbenzene, heteroauxin, benzene and derivative thereof, organic arsenic preparation, organic mercurials, sulfa drug, reach other vegetable alkaloid not following 200 kinds [5～9].Induced mutation rate was very low when above several method used separately, was generally a few percent.
It below is the list of references that background technology is being quoted
[1] Wang Ming vegetable thremmatology (second edition) [M] Beijing: agriculture publishing house, 1990:150～160
[2] Liu Wenge does not have the northern gardening of seed watermelon with few seeds progress [J], 1999, (2): 35～37
[3]Maynard DN.Watermelon cultivars in the United States in2000[J].Cucurbit Coop Rrt，2000，23，51～53
[4] Liu Wenge realizes that watermelon does not have strategy [J] the subject survey China watermelon and muskmelon 2003 (6): 17～20 of seed
[5] Liu Wenge, ploidy operation and ploidy in the breeding of Wang Ming watermelon and muskmelon are identified [J] fruit tree journal, 2,002 19 (2)～13
[6] the big Tianjin of ploidy operation [J] agricultural science 2002 (9) 30～34 in the melon genetic breeding of Han Yike Du triumph
[7] the firm Song Yun vegetables of Qiao Yong polyploid breeding Review Study [J] the Changjiang river vegetables, academic monograph 2,002 8～10
[8] Liu Wenge, Wang Ming vegetable crop multiploid induction progress [J] the Changjiang river vegetables 2003 (01) 29～31
[9] Wang Zhimin, application [J] the biology magazine 2004 (12) 35～38 of ox justice multiploid induction on vegetable breeding
Summary of the invention
At above-mentioned problems of the prior art with not enough, the object of the present invention is to provide a kind of efficiently, the method for seed selection of watermelon polyploid fast.
Realize that foregoing invention purpose technology is a kind of method that improves seed selection of watermelon polyploid, specifically comprise the following steps:
The dliploid seed is soaked seed in normal-temperature water after scalding seed soaking through temperature behind 6～8h, again behind 0.05%～0.1% colchicine seed soaking, 16～20h, wrap the back with wet towel and in incubator, carry out vernalization 48～72h under 28-32 ℃, be seeded in after showing money or valuables one carries unintentionally in the dish of cave, matrix can be selected peat for use, vermiculite, seedling mediums such as perlite, in the greenhouse, grow seedlings, grow to a leaf wholeheartedly or two leaves wholeheartedly the time the watermelon seedling, seedling placed 12 ℃～18 ℃ artificial intelligence incubator, after seedling extractd growing point, carry out colchicine (concentration 0.2%～0.4%) and drip the seedling processing, drip seedling once a day, 12～16h/ is big in illumination, intensity of illumination is to handle 4～6 fates under the 3000lx condition, carries out multiploid induction, treats after seedling grows new growing point seedling to be carried out the polyploid evaluation, with the field planting of polyploid seedling, the polyploid seed is preserved in the training pollination of keeping down the vines of a creeping plant.
Compared with prior art, the invention has the advantages that:
Patent of the present invention has been carried out multifactor integrated treatment with watermelon dliploid seedling first, not only also the watermelon seedling has been carried out certain low temperature treatment simultaneously to having adopted conventional colchicine seedling drop method to handle seedling, handle with pinching, improved the induced mutation rate of watermelon polyploid under the condition of living body greatly.
Embodiment
Below the concrete test example and the embodiment that provide by the inventor further specify effect of the present invention and concrete seed selection of watermelon polyploid method.
Experimental example 1
1 materials and methods
1.1 experiment material
For trying 3 diploid watermelon germplasms:
Red little jade (early-maturing variety): the fruit subcircular, the heavy 3kg of single melon, the light green apparent dark green slice band of pericarp background color, the pericarp crisp fritter, the flesh look red, and quality is husky thin, and the center contains sugar more than 11%, nearly skin zone 8%, seed is little and many.
Capital father-2 (medium variety): the fruit subcircular, the heavy 5kg of single melon, the pericarp background color is light green show dark green in wide band, pericarp is thin and crisp, the flesh toner is red, quality is husky thin, the center contains sugar more than 11%, nearly skin zone 8%, seed is medium and many.
M08 (late variety): the fruit subcircular, the heavy 7kg of single melon, the light green apparent dark green wide band of pericarp background color, pericarp is thin and crisp, and the flesh toner is red, and quality sand is carefully crisp slightly, and the center contains sugar more than 11%, nearly skin zone 8%, seed is medium and many.
1.2 material is cultivated and experiment is handled
1.2.1 the cultivation of material
Seed behind 0.1% colchicine seed soaking 18h, carries out being seeded in the dish of cave after the vernalization in incubator behind normal temperature seed soaking 6～8h again, and matrix is 3: 1 peat and vermiculite, grows seedlings in the greenhouse.
1.2.2 experiment is handled
Different germ plasm resource watermelon seedling are carried out the processing of the integrated treatment in temperature, the pinching of different seedling attitude, colchicine solution concentration, processing time etc.Design the test of four factors, three horizontal quadratures as table 1.
Table 1 watermelon four factors three horizontal quadrature mutagenesis testing tables
| Handle | Colchicine concentration (%) | Treatment temperature | Seedling attitude (several leaves wholeheartedly) pinching | Handle fate |
| 1 2 3 4 5 6 7 8 | 0.20 0.20 0.20 0.30 0.30 0.30 0.40 0.40 | 12 15 18 12 15 18 12 15 | 1 2 3 2 3 1 3 1 | 4 5 6 6 4 5 5 6 |
| 9 | 0.40 | 18 | 2 | 4 |
Annotate: nine processing are established in experiment altogether
When the watermelon seedling grows to suitable seedling attitude, seedling is placed the artificial intelligence incubator that is adjusted to suitable low temperature, seedling is extractd growing point, and carry out colchicine and drip seedling, every afternoon, 18:00 dripped seedling, handle certain fate, temperature constant during the processing, illumination is 3000lx for 7:0-18:00 illumination.
2 experimental results and analysis
Design four factors, three horizontal quadratures test result such as table 2:
Table 2 watermelon orthogonal experiment is handled overall result
| Kind | Handle | Total strain number (strain) | Survive the strain number | Survival rate (%) | The variant number | Induced mutation rate (%) |
| Red little beautiful capital father-2 | 1 2 3 4 5 6 7 8 9 ck1 ck2 1 2 | 75 70 51 68 61 70 51 53 63 46 48 60 66 | 72 67 49 65 59 68 50 51 62 44 46 52 62 | 96.00 95.71 96.08 95.59 96.72 97.14 98.04 96.23 98.41 95.65 95.83 86.67 93.94 | 4 2 9 8 6 13 13 8 5 0 0 3 9 | 5.33 2.86 17.65 11.76 9.84 18.57 25.49 15.09 7.94 0.00 0.00 5.00 13.64 |
| m08 | 3 4 5 6 7 8 9 ck1 ck2 1 2 3 4 5 6 7 8 9 ck1 ck2 | 58 72 58 61 66 72 69 50 50 50 50 50 50 50 48 50 50 48 45 50 | 56 63 57 58 62 65 66 48 49 45 48 49 49 50 46 48 44 45 44 49 | 96.55 87.50 98.28 95.08 93.94 90.28 95.65 96.00 98.00 90.00 96.00 98.00 98.00 100.00 95.83 96.00 88.00 93.75 97.78 98.00 | 5 16 1 7 3 13 8 0 0 12 23 20 21 15 17 19 10 14 12 0 | 8.62 22.22 1.72 11.48 4.55 18.06 11.59 0.00 0.00 24.00 46.00 40.00 42.00 30.00 35.42 38.00 20.00 29.17 26.67 0.00 |
As can be seen from the above table:
2.1 the survival rate after the mutagenesis
As can be seen from Table 2: seedling is through colchicine, and the survival rate of its seedling is mostly more than 85% after pinching and the low temperature treatment, and most of more than 90%, processing 5 survival rates of M08 are up to 100%.Though the seedling percent after the survival rate of seedling all than higher, is handled is lower than not having treated survival rate, and to same germplasm, treatment temperature is low more, seedling is more little, and the survival rate of seedling is also low more, and the survival rate of the same processing of different germplasms seedling is also different.
2.2 the aberration rate interpretation of result after the orthogonal processing
2.2.1 induced mutation rate interpretation of result in nine processing of red little jade:
The red little beautiful induced mutation rate result of table 3
| Handle | 7 | 6 | 3 | 8 | 4 | 5 | 9 | 1 | 2 |
| Average | 25.49 | 18.57 | 17.65 | 15.09 | 11.76 | 9.84 | 7.94 | 5.33 | 2.86 |
As can be seen from the above table: the best processing of red little jade is to handle 7: promptly red little jade is that 0.4% treatment temperature is 12 ℃ of pinching during at 3 leaves, 1 heart in colchicine concentration, the effect of mutagenesis was preferably 25.49% when the processing time was 5 days, next is exactly to handle 6: red little jade is that 0.3% treatment temperature is 18 ℃ of pinching during at 1 leaf, 1 heart in colchicine concentration, treatment effect was also relatively good when the processing time was 5 days is: 18.57%, thereby can select to handle 7 and 6 processing as the inducing watermelon polyploid to the red little jade of germplasm.
2.2.2 capital father-2 induced mutation rate interpretation of result
Table 4 capital father-2 induced mutation rate result
| Handle | 4 | 8 | 2 | 9 | 6 | 3 | 1 | 7 | 5 |
| Average | 22.22 | 18.06 | 13.64 | 11.59 | 11.48 | 8.62 | 5 | 4.55 | 1.72 |
As can be seen from the above table: the best processing of capital father-2 is to handle 4: promptly germplasm capital father-2 is that 0.3% treatment temperature is 12 ℃ of pinching during at 2 leaves, 1 heart in colchicine concentration, the effect of mutagenesis was preferably 22.22% when the processing time was 6 days, next is exactly to handle 8: be that 0.4% treatment temperature is 15 ℃ of pinching during at 1 leaf, 1 heart in colchicine concentration promptly, treatment effect was also relatively good when the processing time was 6 days, be 18.06%, thereby can adopt processing 4 and handle 8 as the processing preferably of inducing capital father-2 multiploid induction concerning germplasm capital father-2.
2.2.3M08 induced mutation rate interpretation of result in nine processing:
Table 5M08 induced mutation rate result
| Handle | 2 | 4 | 3 | 7 | 6 | 5 | 9 | 1 | 8 |
| Average | 46 | 42 | 40 | 38 | 35.42 | 30 | 29.17 | 24 | 20 |
As can be seen from the above table: the best processing of M08 is to handle 2: promptly M08 is that 0.2% treatment temperature is 15 ℃ of pinching during at 2 leaves, 1 heart in colchicine concentration, the effect of mutagenesis was preferably 46% when the processing time was 5 days, next is exactly to handle 4: be that 0.3% treatment temperature is 12 ℃ of pinching during at 2 leaves, 1 heart in colchicine concentration promptly, processing time is 6 days to be that treatment effect is 42% also than higher, thereby can adopt concerning germplasm M08 and handle 2 and handle 4 as the processing preferably of inducing the multiploid induction of M08.
2.3 induce the result to carry out quadrature analysis to above three germplasm watermelon polyploids
The red little beautiful orthogonal experiment quadrature analysis table of table 6
| The factor | Minimum | Maximum | Extreme difference R | Adjust R ' |
| The autumn waters--limid eyes concentration-temperature seedling attitude time | 8.61333 9.26333 7.52000 7.70333 | 16.1733 14.7200 17.6600 15.6400 | 7.5600 5.4567 10.1400 7.9367 | 6.80904 4.91464 9.13276 7.14829 |
| Table 7 capital father-2 orthogonal experiment quadrature analysis table | ||||
| The factor | Minimum | Maximum | Extreme difference R | Adjust R ' |
| Autumn waters--limid eyes concentration | 9.08667 | 11.8067 | 2.7200 | 2.44981 |
| The temperature seedling attitude time | 10.56333 4.96333 6.10333 | 11.1400 15.8167 16.3000 | 0.5767 10.8533 10.1967 | 0.51938 9.77523 9.18380 |
Table 8M08 quadrature analysis table
| The factor | Minimum | Maximum | Extreme difference R | Adjust R ' |
| The autumn waters--limid eyes concentration-temperature seedling attitude time | 29.05667 32.00000 26.47333 27.72333 | 36.6667 34.8633 39.0567 39.8067 | 7.6100 2.8633 12.5833 12.0833 | 6.85407 2.57891 11.33339 10.88305 |
By quadrature analysis, above three watermelon dliploid germplasms to be induced in the process, four factors magnitude relationship that influences to the mutagenesis result in orthogonal experiment is: seedling attitude (pinching)＞processing time＞colchicine concentration〉treatment temperature.Be variation the having the greatest impact that different seedling attitude pinching is handled to whole induced mutation rate, next knows the different processing times, the change in concentration of colchicine besides, be different low temperature treatment at last, thereby different seedling attitude pinching processing is the key factor that improves induced mutation rate, thereby, in mutagenic processes, to select only seedling attitude to induce processing.
3 sum up and discuss
Drip the inductivity that the seedling processing can significantly improve watermelon polyploid 3.1 as can be seen the watermelon seedling is carried out pinching by orthogonal experiments, M08 reaches as high as 46% to the watermelon germplasm, red little jade and capital father also are greatly improved, and have reached 25.49% and 22.22%.And draw by orthogonal experiment that to handle in different seedling attitude pinching be the key factor that improves the polyploid mutagenesis rate, and secondly be the processing time, the variation of the concentration of colchicine besides, what influence was minimum is different low temperature treatment.
3.2 by whole experiment as can be seen, the different germplasms of watermelon are carried out in the polyploid mutagenesis process, seedling is more little, and treatment temperature is low more, and the processing time is long more, and colchicine concentration is high more during processing, and the survival rate of seedling is low more, but basically all more than 85%.
3.3 by to the induction experiment of watermelon polyploid, can must find out obviously that the pinching that watermelon is carried out different seedling attitudes handles the induced mutation rate that can obviously improve watermelon polyploid, obtain the watermelon polyploid germplasm.Owing to watermelon seedling seedling raise period, seedling processing time and cultivation time have scarcely together, have influenced the survival rate of watermelon seedling to a certain extent in this experiment, thereby will eliminate some Effect of Environmental in the test of multiploid induction afterwards as far as possible.，
The production method of embodiment 1 capital father-2 polyploid seed
Diploid watermelon capital father-2 seed is soaked seed in normal-temperature water behind the 6h scalding the seed soaking back through temperature, again behind 0.08% colchicine seed soaking 18h, wrap 30 ℃ of (following vernalization 48～72h that carry out in incubator afterwards with wet towel, be seeded in after showing money or valuables one carries unintentionally in the dish of cave, seedling medium is peat and vermiculite, in the greenhouse, grow seedlings, grow to different seedling attitude two leaves wholeheartedly the watermelon seedling, seedling placed 12 ℃ artificial intelligence incubator, seedling is extractd growing point after, carry out 0.3% colchicine, drip seedling once a day, at illumination 14h/ days, intensity of illumination was 3000lx, handled six days continuously, carry out multiploid induction, treat after seedling grows new growing point seedling to be carried out the polyploid evaluation, the polyploid mutagenesis rate that obtains capital father-2 is 23.2%, with the field planting of polyploid seedling, the polyploid seed is preserved in the training pollination of keeping down the vines of a creeping plant.
The production method of embodiment 2M08 polyploid seed
Diploid watermelon capital father-2 seed is soaked seed in normal-temperature water behind the 8h scalding the seed soaking back through temperature, again behind 0.05% colchicine seed soaking 18h, wrap 30 ℃ of (following vernalization 48～72h that carry out in incubator afterwards with wet towel, be seeded in after showing money or valuables one carries unintentionally in the dish of cave, seedling medium is peat and vermiculite, in the greenhouse, grow seedlings, grow to different seedling attitude two leaves wholeheartedly the watermelon seedling, seedling placed 15 ℃ artificial intelligence incubator, seedling is extractd growing point after, carry out 0.2% colchicine, drip seedling once a day, at illumination 14h/ days, intensity of illumination was 3000lx, handled five days continuously, carry out multiploid induction, treat after seedling grows new growing point seedling to be carried out the polyploid evaluation, the polyploid mutagenesis rate that obtains M08 is 46%, with the field planting of polyploid seedling, the polyploid seed is preserved in the training pollination of keeping down the vines of a creeping plant.
The production method of the yellow polyploid seed in embodiment 3 seas
The seed that diploid watermelon is extra large yellow No. one is soaked seed in normal-temperature water after scalding seed soaking through temperature behind the 8h; again behind 0.0% colchicine seed soaking 18h; wrap 30 ℃ of (following vernalization 48～72h that carry out in incubator afterwards with wet towel; be seeded in after showing money or valuables one carries unintentionally in the dish of cave; seedling medium is peat and vermiculite; in the greenhouse, grow seedlings; grow to different seedling attitude one leaves wholeheartedly the watermelon seedling, seedling placed 18 ℃ artificial intelligence incubator, seedling is extractd growing point after; carry out 0.3% colchicine; drip seedling once a day, at illumination 14h/ days, intensity of illumination was 3000lx; handled five days continuously; carry out multiploid induction, treat after seedling grows new growing point seedling to be carried out the polyploid evaluation, the yellow polyploid mutagenesis rate in many seas that obtains is 28%; with the field planting of polyploid seedling, the polyploid seed is preserved in the training pollination of keeping down the vines of a creeping plant.
Claims (1)
1. method that improves seed selection of watermelon polyploid, it is characterized in that, the dliploid seed is soaked seed in normal-temperature water after scalding seed soaking through temperature behind 6～8h, again behind 0.05%～0.1% colchicine seed soaking, 16～20h, wrap the back with wet towel and in incubator, carry out vernalization 48～72h under 28-32 ℃, be seeded in after showing money or valuables one carries unintentionally in the dish of cave, matrix can be selected peat for use, vermiculite, seedling mediums such as perlite, in the greenhouse, grow seedlings, grow to a leaf wholeheartedly or two leaves wholeheartedly the time the watermelon seedling, seedling placed 12 ℃～18 ℃ artificial intelligence incubator, after seedling extractd growing point, be that 0.2%～0.4% colchicine drips seedling and handles, drip seedling once a day with concentration, at illumination 12～16h/ days, intensity of illumination is to handle 4～6 fates under the 3000lx condition, carries out multiploid induction, treats after seedling grows new growing point seedling to be carried out the polyploid evaluation, with the field planting of polyploid seedling, the polyploid seed is preserved in the training pollination of keeping down the vines of a creeping plant.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101422098B (en) * | 2008-12-03 | 2011-06-22 | 西北农林科技大学 | Method for increasing germination ratio and seedling ratio of triploid watermelon |
| CN103583356A (en) * | 2013-12-02 | 2014-02-19 | 中国科学院新疆生态与地理研究所 | Tetraploid mutagenesis method for cantaloupe |
| CN106106142A (en) * | 2016-06-27 | 2016-11-16 | 广州中医药大学 | A kind of method inducing Herba Andrographis polyploid |
| CN112586348A (en) * | 2020-09-15 | 2021-04-02 | 中国农业科学院郑州果树研究所 | Watermelon tetraploid propagation method and application thereof |
| CN112715355A (en) * | 2020-12-29 | 2021-04-30 | 河南省锄禾园林草业服务有限公司 | Safe and efficient mutation method for bluegrass |
| CN113632728A (en) * | 2021-03-30 | 2021-11-12 | 安徽省农业科学院园艺研究所 | Watermelon polyploid induction and identification method |
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| US10595479B2 (en) * | 2014-04-24 | 2020-03-24 | Purdue Research Foundation | Induced mutagenesis |
Family Cites Families (1)
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| CN1498528A (en) * | 2002-11-06 | 2004-05-26 | 天津科润农业科技股份有限公司蔬菜研 | Method for inducting watermelon bud to tetraploid |
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| CN101422098B (en) * | 2008-12-03 | 2011-06-22 | 西北农林科技大学 | Method for increasing germination ratio and seedling ratio of triploid watermelon |
| CN103583356A (en) * | 2013-12-02 | 2014-02-19 | 中国科学院新疆生态与地理研究所 | Tetraploid mutagenesis method for cantaloupe |
| CN106106142A (en) * | 2016-06-27 | 2016-11-16 | 广州中医药大学 | A kind of method inducing Herba Andrographis polyploid |
| CN112586348A (en) * | 2020-09-15 | 2021-04-02 | 中国农业科学院郑州果树研究所 | Watermelon tetraploid propagation method and application thereof |
| CN112715355A (en) * | 2020-12-29 | 2021-04-30 | 河南省锄禾园林草业服务有限公司 | Safe and efficient mutation method for bluegrass |
| CN113632728A (en) * | 2021-03-30 | 2021-11-12 | 安徽省农业科学院园艺研究所 | Watermelon polyploid induction and identification method |
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