CN113854149A - Method for culturing callus by using wheat microspore - Google Patents
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- CN113854149A CN113854149A CN202111219442.1A CN202111219442A CN113854149A CN 113854149 A CN113854149 A CN 113854149A CN 202111219442 A CN202111219442 A CN 202111219442A CN 113854149 A CN113854149 A CN 113854149A
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- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 35
- 241000209140 Triticum Species 0.000 title claims abstract description 31
- 235000021307 Triticum Nutrition 0.000 title claims abstract description 31
- 238000000034 method Methods 0.000 title claims abstract description 25
- 238000012258 culturing Methods 0.000 title claims abstract description 17
- 239000000463 material Substances 0.000 claims abstract description 27
- 230000006698 induction Effects 0.000 claims abstract description 13
- 239000001963 growth medium Substances 0.000 claims abstract description 12
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- 210000005069 ears Anatomy 0.000 claims description 15
- 239000002609 medium Substances 0.000 claims description 9
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 5
- 239000005631 2,4-Dichlorophenoxyacetic acid Substances 0.000 claims description 5
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 5
- 239000005018 casein Substances 0.000 claims description 5
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 claims description 5
- 235000021240 caseins Nutrition 0.000 claims description 5
- 150000002505 iron Chemical class 0.000 claims description 5
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- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 4
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims description 4
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 2
- 229930195725 Mannitol Natural products 0.000 claims description 2
- 239000002253 acid Substances 0.000 claims description 2
- 239000001110 calcium chloride Substances 0.000 claims description 2
- 229910001628 calcium chloride Inorganic materials 0.000 claims description 2
- 239000000594 mannitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 claims description 2
- CLGDMXUIKIBCOA-XHSDSOJGSA-N C1=CC=CC=2[C@@]34CCCC[C@H]3[C@@H](CC12)N(CC4)S(=O)(=O)O Chemical compound C1=CC=CC=2[C@@]34CCCC[C@H]3[C@@H](CC12)N(CC4)S(=O)(=O)O CLGDMXUIKIBCOA-XHSDSOJGSA-N 0.000 claims 1
- 241001024327 Oenanthe <Aves> Species 0.000 claims 1
- 238000006460 hydrolysis reaction Methods 0.000 claims 1
- 238000005286 illumination Methods 0.000 claims 1
- 230000023409 microsporogenesis Effects 0.000 abstract description 5
- 239000002699 waste material Substances 0.000 abstract description 4
- 238000012216 screening Methods 0.000 abstract 1
- 241000196324 Embryophyta Species 0.000 description 4
- FAIXYKHYOGVFKA-UHFFFAOYSA-N Kinetin Natural products N=1C=NC=2N=CNC=2C=1N(C)C1=CC=CO1 FAIXYKHYOGVFKA-UHFFFAOYSA-N 0.000 description 4
- 238000009395 breeding Methods 0.000 description 4
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- 238000011069 regeneration method Methods 0.000 description 4
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- QANMHLXAZMSUEX-UHFFFAOYSA-N kinetin Chemical compound N=1C=NC=2N=CNC=2C=1NCC1=CC=CO1 QANMHLXAZMSUEX-UHFFFAOYSA-N 0.000 description 3
- 230000001954 sterilising effect Effects 0.000 description 3
- IAKHMKGGTNLKSZ-INIZCTEOSA-N (S)-colchicine Chemical compound C1([C@@H](NC(C)=O)CC2)=CC(=O)C(OC)=CC=C1C1=C2C=C(OC)C(OC)=C1OC IAKHMKGGTNLKSZ-INIZCTEOSA-N 0.000 description 2
- OVSKIKFHRZPJSS-DOMIDYPGSA-N 2-(2,4-dichlorophenoxy)acetic acid Chemical compound OC(=O)[14CH2]OC1=CC=C(Cl)C=C1Cl OVSKIKFHRZPJSS-DOMIDYPGSA-N 0.000 description 2
- 229960005181 morphine Drugs 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
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- HXKWSTRRCHTUEC-UHFFFAOYSA-N 2,4-Dichlorophenoxyaceticacid Chemical compound OC(=O)C(Cl)OC1=CC=C(Cl)C=C1 HXKWSTRRCHTUEC-UHFFFAOYSA-N 0.000 description 1
- 240000005979 Hordeum vulgare Species 0.000 description 1
- 235000007340 Hordeum vulgare Nutrition 0.000 description 1
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- 230000011712 cell development Effects 0.000 description 1
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- 238000012252 genetic analysis Methods 0.000 description 1
- 210000004602 germ cell Anatomy 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
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- 210000000130 stem cell Anatomy 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
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- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
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Images
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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Biotechnology (AREA)
- Cell Biology (AREA)
- Developmental Biology & Embryology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
A method for culturing callus by using wheat microspore comprises screening according to diameter of young ear of wheat when obtaining materials, selecting young ear of wheat with diameter of 0.65-0.74cm, and treating the selected young ear at low temperature; inoculating anther, collecting microspore after rotary cutting, and pretreating the collected microspore in proper solution and temperature; purifying the pretreated microspore, culturing in an induction culture medium, and inducing callus. According to the method, the wheat young ear diameter is used for quantitative material taking, the efficiency and universality of material taking are improved, the operation is simple, the material damage and waste caused by the need of anther color observation or microspore development period observation are effectively avoided, a large number of callus tissues are obtained after culture, the problem that the material cannot be taken for microspore culture due to the requirement on technical equipment can be solved, and the productivity is greatly improved.
Description
Technical Field
The invention belongs to the technical field of crop in-vitro culture, and particularly relates to a method for culturing callus by using wheat microspores.
Background
The microspore is a stem cell of a plant and has strong plasticity and regeneration capacity; meanwhile, the microspore is the germ cell of the plant and has the characteristics of single cell and haploid. By utilizing the characteristics of the microspores, theoretical basic researches such as plant germ cell development, genetic analysis, molecular markers and the like can be carried out; the method can also be applied to haploid breeding, and breeding application research is carried out by one-time homozygosis, accelerating the breeding process and improving the breeding efficiency.
Obtaining a material with a proper microspore development period is a prerequisite for developing free microspore culture. At present, the wheat microspore culture is mainly obtained by adopting a microscopic examination judgment method or an anther color judgment method, and the two methods have higher requirements on laboratory instruments and technical personnel and cannot be universally used by people.
The method for measuring the flag leaf distance is adopted on barley for measurement, but the method for measuring the flag leaf distance cannot be applied to wheat materials, because the flag leaves are extracted when materials suitable for wheat microspore isolated culture are obtained.
Disclosure of Invention
The invention aims to provide a method for culturing callus by utilizing wheat microspores, which is characterized in that the diameter of young ears of wheat is used for quantitatively taking materials, so that the efficiency and universality of material taking are improved, the operation is simple, the material damage and waste caused by the need of anther color observation or microspore development period observation are effectively avoided, the problem that materials cannot be taken for microspore culture due to the requirement on technical equipment is solved, and the productivity is greatly improved.
In order to achieve the purpose, the invention provides the following technical scheme:
a method for culturing callus by using wheat microspore comprises the following steps:
1) taking materials
Taking young ears of wheat drawn from flag leaves, only keeping one leaf, and selecting young ears with the diameter of 0.65-0.74 cm;
2) pretreatment of young ears
Keeping the length of the base part of the selected young ear to be 1.0-2.0cm, cutting off leaves, and placing in a fresh-keeping bag for low-temperature treatment at 4-6 ℃ for 12-21 days;
3) free microspore
Inoculating anther, rotary-cutting at high speed, dissociating and collecting microspore, and pretreating the collected microspore in the extracting solution for 1-3 days at 24-28 deg.C;
wherein the extractive solution contains mannitol 55-65 g.L-1Adding 1-1.2 g.L-1CaCl2、0.9-1.0g·L- 1N-morphinoethanesulfonic acid and 5-15 mg. L-1Colchicine, pH 5.6-6.0;
4) microspore culture and callus induction
And purifying the pretreated microspore, culturing in an induction culture medium, and inducing callus.
Preferably, in step 3), the spin-cut speed is 10000--1Rotary-cut 2-3 times for 10-15 seconds each time.
The extract solution contains 60 g.L of mannitol-1Adding 1.1 g.L-1CaCl2、0.976g·L-1N-morphine ethane sulfonic acid (MES) and 10 mg. L-1Colchicine, pH 5.8.
Further, the induction medium is N6The culture medium is basic culture medium, the iron salt is doubled, and 90 g.L is added-1Maltose, 0.5 mg.L-1Kinetin (KT) 2.0 mg. L-12,4-Dichlorophenoxyacetic acid (2,4-Dichlorophenoxyacetic acid, 2,4-D), 0.976 g.L-1MES、400mg·L-1Hydrolyzed casein and 1600 mg.L-1Glutamine, PH 5.8.
Preferably, the induction medium takes N6 medium as a basic medium, the iron salt is doubled, and 90 g.L < -1 > of maltose, 0.5 mg.L < -1 > of kinetin, 2.0 mg.L < -1 > of 2,4-dichlorophenoxyacetic acid, 0.976 g.L < -1 > of MES, 400 mg.L < -1 > of hydrolyzed casein, 1600 mg.L < -1 > of glutamine and 5.8 of PH are added.
Preferably, the variety of wheat is Alondra.
According to the method, the diameter of the young ear refers to the diameter of the thickest ear, materials are obtained through the diameter of the young ear of the wheat on the premise that the diameter of the young ear of the wheat is strongly related to the development period of microspores, the efficiency and universality of the materials are improved, the operation is simple, material damage and waste caused by anther color observation or microspore development period observation are effectively avoided, the problem that materials cannot be obtained for microspore culture due to the requirement on technical equipment can be solved, when the diameter of the young ear of the wheat is determined, the development period of the microspores is in the middle-late stage of mononucleate, the method is suitable for callus induction, the microspores are subjected to star-shaped division during culture, and more callus tissues can be formed.
Compared with the prior art, the invention has the following beneficial effects:
according to the method, materials are taken according to the diameter of the young ear of the wheat, the requirements on people and equipment are reduced, accurate quantification can be achieved, the efficiency and universality of material taking are improved, the operation is simple, material damage and waste caused by the need of anther color observation or microspore development period observation are effectively avoided, and the productivity is greatly improved.
When young ears with the diameter of 0.65-0.74cm are selected, the development period of the microspores is in the middle-late mononuclear stage, the microspores are subjected to star-shaped division during culture, more callus tissues can be formed, 77.2 regenerated plants can be differentiated from 100mg of the callus tissues, and a foundation is laid for further optimizing green seedling regeneration in the later stage.
Drawings
Figure 1 shows young ears of material Alondra of different diameters.
FIG. 2 shows the callus production of young ears of different diameters in the example of the present invention.
FIG. 3 shows the callus cultured by microspore with different young ear diameters according to the present invention, wherein the young ear diameters are 0.5, 0.6, 0.7 and 0.8cm from left to right.
Detailed Description
The present invention is further illustrated by the following specific examples.
Example a method for obtaining materials by using wheat microspores to culture callus
1. Material
The wheat material Alondra is planted in a phytotron by a pot and is illuminated by 25 mu mol.m-2·s-1，12h·d-1Humidity 70%, temperature 22 ℃ under light and 18 ℃ in darkness.
2. Taking wheat materials
Taking young ears of flag leaves, only keeping one leaf, and dividing into 5 types according to the diameter of the thickest part of the ear, which are 0.35-0.44cm, 0.45-0.54cm, 0.55-0.64cm, 0.65-0.74cm and 0.75-0.84cm respectively, by measuring with vernier caliper.
Fig. 1 shows young ears of the material Alondra with different diameters, and the diameters of the young ears are 0.8, 0.7, 0.6, 0.5 and 0.4cm from left to right.
3. Pretreatment of young ears
Keeping the length of the base part of the young ear to be 1.5cm, cutting off leaves, putting into a freshness protection bag, and putting into a refrigerator with the temperature of 5 ℃ for low-temperature pretreatment for 12-21 days.
4. Collecting microspores
Sterilizing with 10% disinfectant for 10min before inoculation on a sterile super-clean workbench, washing with sterile water for 4-5 times, connecting 5 anthers of ears with each test tube, adding 12ml of the extractive solution, and treating in a refrigerator at 5 deg.C for 2 days.
Wherein the extractive solution contains mannitol 60 g.L-1Adding CaCl2 1.1g·L-1N-morphine ethane sulfonic acid (MES) 0.976 g.L-1And colchicine 10 mg. L-1pH 5.8, filtering and sterilizing.
After the pretreatment of the anther, the anther is cut off by the high-speed disperser on the super clean bench in an overspeed way, the cutting speed is 10000--1Rotary cutting for 2-3 times, each time for 10-15 seconds;
filtering the rotary-cut suspension with 150 mesh screen, and filtering the filtrate at 700 r.min-1Centrifuging at low speed for 5min, repeating for 3 times, collecting microspore, and pretreating the collected microspore with extractive solution at 25 deg.C in dark for 2 days.
5. Microspore culture and callus induction
The pretreated microspores were purified with 21% maltose, washed 1 time with induction medium before cultivation, and then microspore density was adjusted to 1.0X 105each.mL-11.0mL of microspore suspension was inoculated into a petri dish (35 mm. times.12 mm) and 3 replicates were sealed with a sealing membrane and cultured in the dark at 25 ℃ for 22 days to form callus.
Wherein the induction medium is N6The culture medium is a basic culture medium, the iron salt is doubled, and 90 g.L of maltose is added-10.5 mg. L of Kinetin (KT)-12.0 mg. L of 2,4-Dichlorophenoxyacetic acid (2, 4-dichlorphenoxyacetic acid, 2,4-D)-1、MES0.976 g·L-1Hydrolyzed casein 400 mg. L-1And glutamine 1600 mg.L-1And (5) filtering and sterilizing at the pH of 5.8.
The results of the test were analyzed using Microsoft Excel 2010, DPS v version 7.05 data processing system.
Callus yield: transferring the callus mass to differentiation medium when culturing microspore for 22d, sucking liquid culture medium out of culture dish, weighing with electronic balance, and adding mg dish-1Represents; after microspores with different scion diameters are cultured, the obtained healsThe yield of injured tissue is shown in FIGS. 1-2.
As can be seen from FIG. 2, when the diameter of young ear is 0.7cm, the callus yield is the highest, 39.06mg per dish-1And there was a significant difference in callus yield from the other groups.
When the diameter of the young ear is 0.4cm, the microspore is not viable during purification, and the culture is not continued, as can be seen from figure 3, the regeneration capacity of the callus obtained by the culture of different young ear diameters from left to right is different, wherein the regeneration capacity of the obtained callus is strongest when the diameter of the young ear is 0.7cm, and 77.2 regenerated plants can be differentiated from 100mg of callus.
Claims (7)
1. A method for culturing callus by using wheat microspore comprises the following steps:
1) taking materials
Taking young ears of wheat drawn from flag leaves, only keeping one leaf, and selecting young ears with the diameter of 0.65-0.74 cm;
2) pretreatment of young ears
Keeping the length of the base part of the selected young ear to be 1.0-2.0cm, cutting off leaves, and placing in a fresh-keeping bag for low-temperature treatment at 4-6 ℃ for 12-21 days;
3) collecting microspores
Inoculating anther, rotary-cutting at high speed, dissociating and collecting microspore, and pretreating the collected microspore in the extracting solution for 1-3 days at 24-28 deg.C;
wherein the extractive solution contains mannitol 55-65 g.L-1Adding 1-1.2 g.L-1CaCl2、0.9-1.0g·L-1N-morphinoethanesulfonic acid and 5-15 mg. L-1Colchicine, pH 5.6-6.0;
4) microspore culture and callus induction
And purifying the pretreated microspore, culturing in an induction culture medium, and inducing callus.
2. The method for culturing callus with wheat microspore according to claim 1, wherein the rotary cutting speed in the step 3) is 10000-30000 r-min-1Rotary-cut 2-3 times for 10-15 seconds each time.
3. The method for culturing callus using wheat microspore according to claim 1, wherein the extract solution comprises: mannitol 60 g.L-1、CaCl2 1.1g·L-10.976 g.L of N-morphinanesulfonic acid-1Colchicine 10 mg. L-1，pH 5.8。
4. The method for culturing callus on wheat microspore according to claim 1, wherein in step 4), the induction medium is N6The culture medium is basic culture medium, iron salt is doubled, and maltose 75-105 g.L is added-10.4-0.6 mg.L of kinetin-12,4-Dichlorophenoxyacetic acid 1.0-3.0 mg.L-1、MES 0.9-1.0g·L-1Hydrolysis casein 350-450 mg.L-1And glutamine 1200-2000 mg.L-1，PH 5.6-6.0。
5. The method for culturing callus on wheat microspore according to claim 1, wherein in step 4), the induction medium is N6The culture medium is a basic culture medium, the iron salt is doubled, and 90 g.L of maltose is added-10.5 mg. L kinetin-12.0 mg.L of 2,4-dichlorophenoxyacetic acid-1、MES0.976 g·L-1Hydrolyzed casein 400 mg. L-1And glutamine 1600 mg.L-1，PH 5.8。
6. The method for culturing callus on wheat microspore according to claim 1, wherein the variety of wheat is Alondra.
7. The method for culturing callus by using wheat microspores according to claim 1, wherein the young wheat ears in step 1) are from a phytotron planted in a pot, and the culture conditions are as follows: illumination of 25. mu. mol. m-2·s-1，12h·d-1Humidity 70%, temperature 22 deg.C in light and 18 deg.C in dark.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115281185A (en) * | 2022-07-05 | 2022-11-04 | 河北省农林科学院生物技术与食品科学研究所 | Method for prolonging preservation time of wheat anther |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2054607A1 (en) * | 1990-11-01 | 1992-05-02 | Kazutoshi Ito | Method for preparing transformed plant |
| JP2008212048A (en) * | 2007-03-02 | 2008-09-18 | National Agriculture & Food Research Organization | Method for improving expression efficiency of target gene introduced into wheat |
| CN109105258A (en) * | 2018-08-15 | 2019-01-01 | 上海市农业科学院 | A kind of cultural method of barley microspore |
| CN110317772A (en) * | 2019-07-24 | 2019-10-11 | 上海市农业科学院 | A kind of method of drawing material of barley microspore |
| CN111454874A (en) * | 2020-03-06 | 2020-07-28 | 河南农业大学 | Method for separating pollen of mononuclear, binuclear and mature corn and construction and application of pollen development stage judgment model |
-
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Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2054607A1 (en) * | 1990-11-01 | 1992-05-02 | Kazutoshi Ito | Method for preparing transformed plant |
| JP2008212048A (en) * | 2007-03-02 | 2008-09-18 | National Agriculture & Food Research Organization | Method for improving expression efficiency of target gene introduced into wheat |
| CN109105258A (en) * | 2018-08-15 | 2019-01-01 | 上海市农业科学院 | A kind of cultural method of barley microspore |
| CN110317772A (en) * | 2019-07-24 | 2019-10-11 | 上海市农业科学院 | A kind of method of drawing material of barley microspore |
| CN111454874A (en) * | 2020-03-06 | 2020-07-28 | 河南农业大学 | Method for separating pollen of mononuclear, binuclear and mature corn and construction and application of pollen development stage judgment model |
Non-Patent Citations (1)
| Title |
|---|
| 杨筱: "小麦成熟胚愈伤组织诱导及分化的研究", 《中国优秀博硕士学位论文全文数据库(硕士)农业科技辑》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115281185A (en) * | 2022-07-05 | 2022-11-04 | 河北省农林科学院生物技术与食品科学研究所 | Method for prolonging preservation time of wheat anther |
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Application publication date: 20211231 |