CN116671436A - Composition for doubling haploid corn and haploid corn doubling method - Google Patents
Composition for doubling haploid corn and haploid corn doubling method Download PDFInfo
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- CN116671436A CN116671436A CN202310866560.4A CN202310866560A CN116671436A CN 116671436 A CN116671436 A CN 116671436A CN 202310866560 A CN202310866560 A CN 202310866560A CN 116671436 A CN116671436 A CN 116671436A
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- haploid
- doubling
- corn
- composition
- haploid corn
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- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 36
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 32
- 235000005822 corn Nutrition 0.000 title claims abstract description 32
- 238000000034 method Methods 0.000 title claims abstract description 23
- 239000000203 mixture Substances 0.000 title claims abstract description 17
- 241000209149 Zea Species 0.000 title 2
- 240000008042 Zea mays Species 0.000 claims abstract description 34
- IAZDPXIOMUYVGZ-UHFFFAOYSA-N Dimethylsulphoxide Chemical compound CS(C)=O IAZDPXIOMUYVGZ-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000005602 Propyzamide Substances 0.000 claims abstract description 11
- PHNUZKMIPFFYSO-UHFFFAOYSA-N propyzamide Chemical compound C#CC(C)(C)NC(=O)C1=CC(Cl)=CC(Cl)=C1 PHNUZKMIPFFYSO-UHFFFAOYSA-N 0.000 claims abstract description 11
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 claims abstract description 10
- 229920000053 polysorbate 80 Polymers 0.000 claims abstract description 10
- 239000005885 Buprofezin Substances 0.000 claims abstract description 5
- PRLVTUNWOQKEAI-VKAVYKQESA-N buprofezin Chemical compound O=C1N(C(C)C)\C(=N\C(C)(C)C)SCN1C1=CC=CC=C1 PRLVTUNWOQKEAI-VKAVYKQESA-N 0.000 claims abstract description 5
- SNPPWIUOZRMYNY-UHFFFAOYSA-N bupropion Chemical compound CC(C)(C)NC(C)C(=O)C1=CC=CC(Cl)=C1 SNPPWIUOZRMYNY-UHFFFAOYSA-N 0.000 claims abstract description 4
- 229960001058 bupropion Drugs 0.000 claims abstract description 4
- 238000003973 irrigation Methods 0.000 claims description 8
- 230000002262 irrigation Effects 0.000 claims description 8
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 claims description 4
- 235000009973 maize Nutrition 0.000 claims description 4
- 239000002904 solvent Substances 0.000 claims description 4
- 238000009395 breeding Methods 0.000 abstract description 5
- 230000001488 breeding effect Effects 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 description 21
- 210000005069 ears Anatomy 0.000 description 14
- 235000013339 cereals Nutrition 0.000 description 12
- 230000006698 induction Effects 0.000 description 11
- 230000000694 effects Effects 0.000 description 7
- 210000001161 mammalian embryo Anatomy 0.000 description 7
- 239000000463 material Substances 0.000 description 7
- 230000010152 pollination Effects 0.000 description 6
- 238000009331 sowing Methods 0.000 description 5
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 5
- 238000003306 harvesting Methods 0.000 description 3
- 230000001939 inductive effect Effects 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 description 3
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 229920001131 Pulp (paper) Polymers 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- SKOLWUPSYHWYAM-UHFFFAOYSA-N carbonodithioic O,S-acid Chemical compound SC(S)=O SKOLWUPSYHWYAM-UHFFFAOYSA-N 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 238000005553 drilling Methods 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 238000009396 hybridization Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000000394 mitotic effect Effects 0.000 description 1
- 239000003960 organic solvent Substances 0.000 description 1
- 230000035515 penetration Effects 0.000 description 1
- 239000002689 soil Substances 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000009736 wetting Methods 0.000 description 1
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)
Abstract
The invention provides a composition for doubling haploid corn and a method for doubling haploid corn, and relates to the technical field of haploid breeding. Comprising the following steps: propyzamide, dimethyl sulfoxide, tween 80 and buprofezin; the concentration of the propyzamide is 80 mu mol/L; the mass percentage of the dimethyl sulfoxide is 1%; the mass percentage of the Tween 80 is 0.05%; the mass percentage of the bupropion is 0.05%. The composition provided by the invention doubles haploid corn, and the doubling rate can reach more than 15% and up to 45%.
Description
Technical Field
The invention relates to the technical field of haploid breeding, in particular to a composition for doubling haploid corn and a method for doubling haploid corn.
Background
The breeding of the maize inbred line by using the haploid technology has the advantages of short time, high homozygosity and the like. Haploid breeding techniques are simpler in the haploid induction stage, but have problems in the doubling stage, such as too low doubling rates and less practical significance if natural doubling is used.
Disclosure of Invention
In order to solve the problems, the invention provides a composition for doubling haploid corn and a method for doubling haploid corn, which improve doubling effect.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides a haploid corn doubling composition, which takes water as a solvent and comprises the following components: propyzamide, dimethyl sulfoxide, tween 80 and buprofezin;
the concentration of the propyzamide is 80 mu mol/L;
the mass percentage of the dimethyl sulfoxide is 1%;
the mass percentage of the Tween 80 is 0.05%;
the mass percentage of the bupropion is 0.05%.
The invention also provides a method for doubling haploid corn, which comprises the following steps:
when the haploid corn grows to 2-3 unfolding leaves, the haploid corn is dripped with the composition according to the technical scheme.
Preferably, the amount of the dripping core is 1ml.
Preferably, the root irrigation amount is 4ml.
Preferably, the root irrigation includes: the composition was placed in the hole with holes perforated in the ground 2cm from the haploid corn root.
Preferably, the diameter of the hole is 0.5cm and the depth of the hole is 5cm.
The beneficial effects of the invention are as follows:
the composition provided by the invention doubles haploid corn, and the doubling rate can reach more than 15% and up to 45%.
Detailed Description
The invention provides a haploid corn doubling composition, which takes water as a solvent and comprises the following components: propyzamide, dimethyl sulfoxide, tween 80 and buprofezin; the concentration of the propyzamide is 80 mu mol/L; the mass percentage of the dimethyl sulfoxide is 1%; the mass percentage of the Tween 80 is 0.05%; the mass percentage of the bupropion is 0.05%. In the invention, the propyzamide has a doubling effect, the dimethyl sulfoxide is an organic solvent, and the tween 80 and the buprofezin are used as auxiliary reagents, so that the toxicity is low, the penetration and adhesion capability of the propyzamide are enhanced, the absorption effect of haploid corn is increased, and the doubling effect is improved.
The invention also provides a method for doubling haploid corn, which comprises the following steps: when the haploid corn grows to 2-3 unfolding leaves, the haploid corn is dripped with the composition according to the technical scheme. In the present invention, the amount of the center drop is preferably 1ml. In the present invention, the root irrigation amount is preferably 4ml. In the invention, the core dripping and root irrigation are performed simultaneously, the double acting time is increased, the core dripping and root irrigation are used in too small amount and have no effect, and the excessive amount can cause toxicity and even death of seedlings. In the present invention, the root irrigation preferably includes: the composition was placed in the hole with holes perforated in the ground 2cm from the haploid corn root. In the present invention, the diameter of the hole is preferably 0.5cm, and the depth of the hole is preferably 5cm. The method is not particularly limited to the haploid corn, and the corn is a haploid.
The present invention will be described in detail with reference to examples for further illustration of the invention, but they should not be construed as limiting the scope of the invention.
Example 1
(1) Selection of inducible lines
The hybrid seeds between CAU and the self-bred inbred Handan induced line No. 1 are used as the induction materials, (Handan induced line No. 1 is the self-bred line, which is formed by breeding the hybrid offspring of CAU5 and H7, and H7 is the female parent of Handan Feng 79), and the hybrid seeds between the induced lines are used as the induction materials, so that the pollen amount is large, the plant height is large, the male parent is easy to be used, and the large-scale induction is performed.
(2) Inducing haploid kernels
The induced material is used as female parent, the hybrid seeds between the induction lines are used as male parent, and the clusters are obtained by hybridization. The planting modes of the female parent and the male parent are two: first, the female parent and the male parent can be planted independently, the female parent is bagged in the flowering period, and pollination is carried out with the female parent filaments by manually collecting the male parent for pollination so as to obtain haploid induction material ears. Secondly, the planting of the male parent and the female parent adopts the planting mode of maize seed production, the planting is carried out according to the ratio of 1:4 of the male parent to the female parent or other ratios, the male parent is properly planted in about 3 days apart and 3 periods, the male parent is removed in advance in the flowering period, the pollination is carried out in a natural pollination mode, and a large quantity of haploid induction material ears are obtained in the later period.
(3) Selection of haploid kernels
The grains on the clusters of the induction material contained 3 conditions: neither embryo nor endosperm is developed, and the seed is pollen which is a hybrid non-inducing material, and is an ineffective seed. Both the endosperm and embryo of the kernel develop, generally purple black, to induce unsuccessful kernels and ineffective kernels. The endosperm of the seed is colored, the embryo is not colored, and the seed is effective. Then, under the condition of planting the effective seeds, doubling treatment is carried out, and then the doubled normal seeds are obtained.
During the planting in the doubling stage, haploid grains are selected out, and a manual selection mode is adopted. The method is carried out when the seeds are not fully mature and are dried in the air, the water content of the seeds is about 30% in a period of about one week in a normal harvest period, the seeds with the color developed by the embryo are directly observed on the ears, the seeds with the color not developed by the embryo and the color developed by the endosperm are selected, the color developed by the embryo is obvious compared with that of the seeds after the seeds are dried in the air, the seeds are easy to select, the accuracy is high, and the efficiency is several times higher than that of the seeds after the seeds are dried in the air.
(4) Haploid seed sowing
Northern in sowing period suggests summer sowing after 6 months, and the general density can be 15000 plants/mu.
(5) Field management
Non-haploid plants can be removed from the period from sowing to jointing, and the appearance is that the plants are obviously thick and strong, high and large, and the stalks and leaves are purple. Fertilizing and other management are the same as those of the common corn cultivation fields. However, water conservation is guaranteed, strong seedlings are cultivated in the early stage, soil wetting is guaranteed especially in the powder scattering stage and a period of time before the powder scattering stage, and plant powder scattering is facilitated.
(6) Preparation of treatment agent
80umol/L of propyzamide, 1% of dimethyl sulfoxide, 0.05% of Tween 80 and 0.05% of thioketone, and water as a solvent; the effect is to suppress the mitotic phase of germ cells of the immature embryo, thereby doubling the cell chromosome.
(7) Medicament treatment
When corn grows to 2-3 expanded leaves, the prepared treatment agent is used for dripping fully in the leaves of the seedlings; simultaneously, a short rod with the diameter of about 0.5cm is used for drilling a deep hole with the diameter of 5cm towards the bottom direction of the corn at the position about 2cm away from the root of the seedling, and the deep hole is filled with the medicament to finish the treatment, 1 milliliter of the core is dropped, and 4 milliliters of the root is filled.
(8) Pollination
And after sowing for about 40 days, observing the growth condition of haploid plants every day, covering the grown female ears with a sulfuric acid paper bag in time, and observing whether pollen is scattered. The doubled haploid pollen scattering condition is far inferior to that of common corn and diploid inbred lines, and the pollination stage is completed until all plants spend the flowering period.
(9) Harvesting
Each plant has a solid haploid plant and represents a genotype, and the genotypes of seeds on the same plant are completely consistent. It is also interesting to harvest only one grain of the ear. Haploid plant 1200 obtained by induction of PH4CV/HCL645 offspring is co-planted in 2020: the 800 plants treated by the method obtain 130 ears of clusters with more than 3 grains, and the doubling rate is 16.3%; only 200 plants treated by the heart dropping method are used for obtaining 16 ears of clusters exceeding 3 grains, and the multiplying power is 8%; 200 plants without any treatment obtained 3 ears of more than 3 grains, with a multiplying power of 1.5%. And (3) co-planting 2000 haploid plants obtained by PH4 CV/Zheng 58 offspring induction in 2021: 1500 strains treated by the method obtain ears 456 ears with more than 3 grains, and the doubling rate is 12.3%; only 300 plants treated by the heart dropping method are used for obtaining 37 ears of clusters with more than 3 grains, and the multiplying power is 8%; 200 plants without any treatment obtained 8 ears of more than 3 grains, doubling rate 4%. Haploid plants 800 obtained by induction of PH4 CV/Zheng 58 offspring were co-planted in 2022: 600 plants treated by the method obtain clusters 269 ears with more than 3 grains, and the doubling rate is 44.8%; only 100 plants treated by the heart dropping method are used for obtaining 13 ears of clusters with more than 3 grains, and the doubling rate is 13%; 100 plants without any treatment obtained 5 ears of more than 3 grains, doubling rate 5%.
The experiment shows that the method has obvious effect on doubling haploid, and has simple operation and great practical value. The highest 3-grain adding multiplying power of the method is 45%, and the highest adding multiplying power is generally 15%.
Although the foregoing embodiments have been described in some, but not all, embodiments of the invention, it should be understood that other embodiments may be devised in accordance with the present embodiments without departing from the spirit and scope of the invention.
Claims (6)
1. A haploid maize doubling composition, characterized in that the composition is water-in-solvent, comprising: propyzamide, dimethyl sulfoxide, tween 80 and buprofezin;
the concentration of the propyzamide is 80 mu mol/L;
the mass percentage of the dimethyl sulfoxide is 1%;
the mass percentage of the Tween 80 is 0.05%;
the mass percentage of the bupropion is 0.05%.
2. A method of doubling haploid maize, comprising the steps of:
dropping the haploid corn and rooting the composition of claim 1 when the haploid corn grows to 2-3 expanded leaves.
3. The method of claim 2, wherein the core is used in an amount of 1ml.
4. The method according to claim 2, wherein the root irrigation amount is 4ml.
5. The method of claim 2 or 4, wherein the root irrigation comprises: the composition was placed in the hole with holes perforated in the ground 2cm from the haploid corn root.
6. The method of claim 5, wherein the holes have a diameter of 0.5cm and a depth of 5cm.
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| Application Number | Priority Date | Filing Date | Title |
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| CN202310866560.4A CN116671436B (en) | 2023-07-14 | 2023-07-14 | Composition for doubling haploid corn and haploid corn doubling method |
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| CN202310866560.4A CN116671436B (en) | 2023-07-14 | 2023-07-14 | Composition for doubling haploid corn and haploid corn doubling method |
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| CN116671436B CN116671436B (en) | 2024-03-12 |
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Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100169999A1 (en) * | 2008-12-29 | 2010-07-01 | Dow Agrosciences Llc | Induced chromosome doubling in plants |
| 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 |
| CN103749291A (en) * | 2013-10-21 | 2014-04-30 | 广西壮族自治区农业科学院玉米研究所 | Method for doubling chromosome complement of maize haploid plant |
| CN103828714A (en) * | 2014-03-07 | 2014-06-04 | 毕节市农业科学研究所 | Method for induction and diploidization of different-germplasm haploid of corn in northwest Guizhou |
| US20140259209A1 (en) * | 2013-03-08 | 2014-09-11 | Dow Agrosciences Llc | Root soaking method to produce double haploid inbred lines in corn |
| CN107018895A (en) * | 2016-01-29 | 2017-08-08 | 辽宁省农业科学院玉米研究所 | A kind of abductive approach of tetraploid silage corn |
| US20190014730A1 (en) * | 2015-07-16 | 2019-01-17 | Monsanto Technology Llc | Methods for Creating Doubled Haploid Plants |
-
2023
- 2023-07-14 CN CN202310866560.4A patent/CN116671436B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100169999A1 (en) * | 2008-12-29 | 2010-07-01 | Dow Agrosciences Llc | Induced chromosome doubling in plants |
| 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 |
| US20140259209A1 (en) * | 2013-03-08 | 2014-09-11 | Dow Agrosciences Llc | Root soaking method to produce double haploid inbred lines in corn |
| CN103749291A (en) * | 2013-10-21 | 2014-04-30 | 广西壮族自治区农业科学院玉米研究所 | Method for doubling chromosome complement of maize haploid plant |
| CN103828714A (en) * | 2014-03-07 | 2014-06-04 | 毕节市农业科学研究所 | Method for induction and diploidization of different-germplasm haploid of corn in northwest Guizhou |
| US20190014730A1 (en) * | 2015-07-16 | 2019-01-17 | Monsanto Technology Llc | Methods for Creating Doubled Haploid Plants |
| CN107018895A (en) * | 2016-01-29 | 2017-08-08 | 辽宁省农业科学院玉米研究所 | A kind of abductive approach of tetraploid silage corn |
Non-Patent Citations (2)
| Title |
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| 惠国强等: "不同除草剂对玉米单倍体成熟胚的加倍效果", 《山西农业科学》, vol. 48, no. 08, pages 1189 - 1191 * |
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