CN102550396A - Method for quickly identifying cross-compatibility of rose hybrida - Google Patents
Method for quickly identifying cross-compatibility of rose hybrida Download PDFInfo
- Publication number
- CN102550396A CN102550396A CN2012100408990A CN201210040899A CN102550396A CN 102550396 A CN102550396 A CN 102550396A CN 2012100408990 A CN2012100408990 A CN 2012100408990A CN 201210040899 A CN201210040899 A CN 201210040899A CN 102550396 A CN102550396 A CN 102550396A
- Authority
- CN
- China
- Prior art keywords
- pollen
- sample
- hours
- bagging
- compatibility
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Landscapes
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a method for quickly identifying cross-compatibility of rose hybrida. The method is characterized by comprising the following steps of: collecting and storing pollen, performing castration, performing artificial pollination, cutting a sample to be identified, fixing the cut sample in FAA stationary liquid, transferring the cut sample to a 70 percent ethanol solution after 24 hours, taking out for washing, placing in an 8mol/L NaOH solution, performing water bath at the temperature of 55 to 60 DEGC for 15 to 20 minutes, and then standing at the temperature of 20 to 28 DEGC for 3 to 3.5 hours, soaking into water for at least 1 hour, transferring to a 0.1 percent dye solution of aniline blue, dyeing for 2 to 3 hours, observing stigma callose deposition, pollen germination and growth of pollen tubes by using a fluorescence microscope to identify the cross-compatibility of rose hybrida. By the method, the cross-compatibility of rose hybrida can be identified within 30 to 48 hours; and moreover, the method is simple and easy to operate and can effectively reduce the waste of breeding resources and improve breeding efficiency.
Description
?
Technical field
The present invention relates to the plant reproduction biology field, specifically is a kind of rapid identification method of Chinese rose cross compatibility.
Background technology
Chinese rose (Rosa hybrida) is the big ornamental plants of the first in the world, and up to now, global modern rose cultivars is above 35000.Most modern rose cultivarses all are to cultivate through long-term edge far away, nearly edge hybridization.Along with the enforcement of UPOV pact 1991 texts, crossbreeding will become the only resource of cultivating the Chinese rose new varieties.Research combo parent's cross compatibility is most important to the seed selection modern rose cultivars, and the application study of especially breeding Core Germplasms resource utilizes distant hybridization screening resistant variety.The Chinese rose cross compatibility receives the control of aspects such as hereditary affiliation, gene, ploidy or influences; On the basis of molecular breeding means research breeding resource heredity affiliation, field cross-fertile rate is that cross compatibility the most truly and directly embodies.The cross incompatibility of Chinese rose has following performance: during hybridization pollination, 1) pollen grain can not be sprouted on column cap; 2) though pollen grain can be sprouted on column cap, pollen tube can not pass column cap or can not pass style and arrive blastular; 3) though pollen tube can arrive blastular, the sperm that it discharges can not merge with egg cell and form zygote, finally causes hybridizing shaky.In the conventional hybridization breeding process; Can only whether expand through observation pollination back ovary and identify whether father and mother are originally affine; And this need just can observe about 15-20 after the pollination days usually; So not only waste a large amount of breeding resources, but also reduced breeding efficiency, be unfavorable for the industry development of high-quality Chinese rose flowers.
Summary of the invention
To cross-incompatible kind; It can produce a large amount of calloses in the pollen tube growth process, callose is β-1, the 3-glucan; Often be distributed in the screen casing of higher plant, new cell wall, pollen grain and the pollen tube that forms; After callose dyes with aniline blue, under ultraviolet excitation, can send yellow to yellowish green fluorescence.Thus; After can dyeing the ovary after the pollination with aniline blue, be put under the fluorescence microscope and observe, promptly can be observed the germinating of pollen on column cap, the growth conditions of pollen tube; And callose is in deposition situation of stigma surface etc., and whether can judge this hybrid combination with this affine.
In order to identify combo parent's cross compatibility as early as possible at short notice, confirm that can group gas-mixing hybridization successful, reduce the breeding wasting of resources, improve breeding efficiency, the present invention provides a kind of rapid identification method of Chinese rose cross compatibility.
The present invention is to provide a kind of like this Chinese rose cross compatibility rapid identification method, the collection that it is characterized in that comprising pollen with preserve step, castrate step, artificial pollination step, sample to be identified cut step, it is characterized in that also comprising the following steps:
A, the sample to be identified that will cut are fixed in the FAA fixer, after 24 hours, transfer to mass concentration and are in 70% the ethanolic solution and preserve; Wherein, said FAA fixer is following proportioning:
Mass concentration is 37～40% formaldehyde 5～10 ml
Mass concentration is 70% ethanol 80～90 ml
Glacial acetic acid 5～10 ml;
B, sample that steps A was fixed by conventional water flushing after, place the NaOH solution of 8mol/L, 55～60 ℃ of water-baths 15～20 minutes were taken out, in 20～28 ℃ of held 3～3.5 hours;
C, the sample of step B was soaked 1 hour with running water at least, change running water therebetween 3～4 times, remove most of sodium hydroxide solution, transfer to mass concentration afterwards and be in 0.1% the aniline blue dye liquor and dyeed 2～3 hours;
D, get the dyeing sample of step C, by routine with fluorescence microscope column cap callose deposition situation, the pollen germination state, the pollen tube growth state, thereby identify the Chinese rose cross compatibility.
The glacial acetic acid of said steps A is commercial analysis net product.
The collection of said pollen with the preservation step is: selects the plant of robust growth, selects and treat open flower, and the mark of listing, the maternal castration, bagging will place 20～25 ℃ of ventilations to dry in the shade from the flower pesticide that the male parent flower is taken, and get loose powder, and 4 ℃ of preservations are for use.
Said castration step is: select the flower that will open next day on the maternal plant to castrate, afterwards bagging.At this moment still incomplete development is ripe for stamen, and pollen can not be scattered during castration.Operating time with afternoon 15:00～17:00 for well.
Said artificial pollination step is: in 9:00-10:00 in the morning, take off the bagging in the maternal strain earlier, dip in writing brush and get the pollen that scatters in right amount, smear pollination on maternal column cap, bagging kept 30～48 hours again.
The step that cuts of said sample to be identified is: take off bagging, the style after gynobase downcuts pollination is together with ovary.
Among the step B of the present invention; The purpose of handling style with NaOH is: remove the protoplasm in the style histocyte; The style histocyte blocks to make pollen tube not receive on every side, is easy to be colored and observe, therefore; This step B is a key link of observing germination of pollen tube, directly has influence on the effect of observation.
When observing sample one of following characteristic is arranged, it is not affine then to be accredited as hybridization:
(1) do not see that pollen germination is arranged.
(2) even if a small amount of pollen germination is arranged, promptly stop growing but grow into the style middle and upper part, the pollen tube end expands, or crooked, or distortion, and column cap and pollen tube have a large amount of callose depositions.
When observing sample and pollen germination is arranged and have a small amount of pollen tube to get into ovary, then being accredited as hybridization has compatibility.
The present invention has advantage and effect: adopt such scheme, but within a short period of time, promptly 30～48 hours; Identify the cross compatibility of Chinese rose combination, fundamentally solve in the conventional hybridization breeding, need whether expand through observing pollination back ovary; Whether originally affine, and this observation need wait the pollination back could observe in about 15-20 days if just can identify father and mother, the inventive method is simple; Strong operability, evaluation combo parent's that can be as early as possible cross compatibility confirms that can group gas-mixing hybridization successful; Effectively reduce the breeding wasting of resources, improve breeding efficiency.
Description of drawings
Fig. 1 is the affine sprouting of pollen on column cap of hybridization;
Fig. 2 is the affine sprouting of pollen on column cap of hybridization;
Fig. 3
ForThe pollen tube at style middle part;
Fig. 4 gets into ovary for the affine pollen tube of hybridization;
Fig. 5 is that amplify the part of Fig. 3;
Fig. 6 gets into ovary for the affine pollen tube of hybridization;
Fig. 7 does not have pollen grain on the cross-incompatible column cap;
Fig. 8 is for there being a large amount of callose depositions on the cross-incompatible column cap.
Embodiment
Below in conjunction with embodiment the present invention is done and to further describe.
Embodiment 1
Chinese rose in the present embodiment 1 refer to rose family Rosa (
RosaLinnaeus) cultivar in (cultivar), process the following step:
1) collection of pollen and preservation: select the plant of robust growth, select and treat open flower, and the mark of listing, will place 20 ℃ of ventilations to dry in the shade from the flower pesticide that the male parent flower is taken, the pollen that must scatter, 4 ℃ of preservations, for use;
2) castrate: in 15:00 in afternoon, select the flower that will open next day on the maternal plant to castrate, bagging afterwards, because of the incomplete development maturation still of stamen at this moment, pollen can not be scattered during castration;
3) artificial pollination: in 9:00 in the morning, take off the bagging in the maternal strain earlier, dip in writing brush and get the pollen that scatters in right amount; Smear pollination on maternal column cap; Bagging kept 30 hours again, and on board, write information such as combination title or code name, hybridization date, operator's name exactly;
4) cutting of sample to be identified: take off bagging, the style after gynobase downcuts pollination is together with ovary;
5) sample to be identified that step 4) is cut is fixed in the FAA fixer, after 24 hours, transfers to mass concentration and is in 70% the ethanolic solution and preserve; Wherein this FAA fixer is following proportioning:
Mass concentration is 37% formaldehyde 10 ml
Mass concentration is 70% ethanol 80 ml
Analyze pure glacial acetic acid 10 ml
6) sample of step 5) being fixed by conventional water flushing after, place the NaOH solution of 8mol/L, 55 ℃ of water-baths 15 minutes were taken out, in 20 ℃ of held 3.5 hours;
7) sample of step 6) was soaked 1 hour with running water at least, change running water therebetween 3 times, remove most of sodium hydroxide solution, transferred to mass concentration afterwards and be in 0.1% the aniline blue dye liquor dyeing 2 hours;
8) with the dyeing sample of one piece of step 7) of tweezers picking, be placed on the slide, with filter paper sucking-off dye liquor, drip a last glycerine; Covered is struck the gland slide gently, and style is launched; Be put into slide under the LEICA DM6000 B fluorescence microscope then and observe, observing column cap has pollen germination, like Fig. 1; And there is a small amount of pollen tube to get into ovary when sampling is observed once more after 18 hours,, explains that this Chinese rose hybridization has compatibility like Fig. 4.
Embodiment 2
Chinese rose in the present embodiment 2 refer to rose family Rosa (
RosaLinnaeus) cultivar in (cultivar), process the following step:
1) collection of pollen and preservation: select the plant of robust growth, select and treat open flower, and the mark of listing, will place 25 ℃ of ventilations to dry in the shade from the flower pesticide that the male parent flower is taken, the pollen that must scatter, 4 ℃ of preservations, for use;
2) castrate: in 17:00 in afternoon, select the flower that will open next day on the maternal plant to castrate, bagging afterwards, because of the incomplete development maturation still of stamen at this moment, pollen can not be scattered during castration;
3) artificial pollination: in 10:00 in the morning, take off the bagging in the maternal strain earlier, dip in writing brush and get the pollen that scatters in right amount; Smear pollination on maternal column cap; Bagging kept 48 hours again, and on board, write information such as combination title or code name, hybridization date, operator's name exactly;
4) cutting of sample to be identified: take off bagging, the style after gynobase downcuts pollination is together with ovary;
5) sample to be identified that step 4) is cut is fixed in the FAA fixer, after 24 hours, transfers to mass concentration and is in 70% the ethanolic solution and preserve; Wherein this FAA fixer is following proportioning:
Mass concentration is 40% formaldehyde 5 ml
Mass concentration is 70% ethanol 90 ml
Analyze pure glacial acetic acid 5 ml;
6) sample of step 5) being fixed by conventional water flushing after, place the NaOH solution of 8mol/L, 60 ℃ of water-baths 20 minutes were taken out, in 28 ℃ of held 3 hours;
7) sample of step 6) was soaked 1 hour with running water at least, change running water therebetween 4 times, remove most of sodium hydroxide solution, transferred to mass concentration afterwards and be in 0.1% the aniline blue dye liquor dyeing 3 hours;
8) with the dyeing sample of one piece of step 7) of tweezers picking, be placed on the slide, with filter paper sucking-off dye liquor, drip a last glycerine; Covered is struck the gland slide gently, and style is launched; Be put into slide under the LEICA DM6000 B fluorescence microscope then and observe, observing column cap has pollen germination, like Fig. 2; And there is a small amount of pollen tube to get into ovary,, explains that this Chinese rose hybridization has compatibility like Fig. 6.
Embodiment 3
Chinese rose in the present embodiment 3 refer to rose family Rosa (
RosaLinnaeus) cultivar in (cultivar), process the following step:
1) collection of pollen and preservation: select the plant of robust growth, select and treat open flower, and the mark of listing, will place 22 ℃ of ventilations to dry in the shade from the flower pesticide that the male parent flower is taken, the pollen that must scatter, 4 ℃ of preservations, for use;
2) castrate: in 15:30 in afternoon, select the flower that will open next day on the maternal plant to castrate, bagging afterwards, because of the incomplete development maturation still of stamen at this moment, pollen can not be scattered during castration;
3) artificial pollination: in 9:40 in the morning, take off the bagging in the maternal strain earlier, dip in writing brush and get the pollen that scatters in right amount; Smear pollination on maternal column cap; Bagging kept 38 hours again, and on board, write information such as combination title or code name, hybridization date, operator's name exactly;
4) cutting of sample to be identified: take off bagging, the style after gynobase downcuts pollination is together with ovary;
5) sample to be identified that step 4) is cut is fixed in the FAA fixer, after 24 hours, transfers to mass concentration and is in 70% the ethanolic solution and preserve; Wherein this FAA fixer is following proportioning:
Mass concentration is 38% formaldehyde 7 ml
Mass concentration is 70% ethanol 85 ml
Analyze pure glacial acetic acid 8 ml;
6) sample of step 5) being fixed by conventional water flushing after, place the NaOH solution of 8mol/L, 58 ℃ of water-baths 18 minutes were taken out, in 25 ℃ of held 3 hours;
7) sample of step 6) was soaked 1 hour with running water at least, change running water therebetween 3 times, remove most of sodium hydroxide solution, transferred to mass concentration afterwards and be in 0.1% the aniline blue dye liquor dyeing 2 hours;
8) with the dyeing sample of one piece of step 7) of tweezers picking, be placed on the slide, with filter paper sucking-off dye liquor, drip a last glycerine; Covered is struck the gland slide gently, and style is launched; Be put into slide under the LEICA DM6000 B fluorescence microscope then and observe, observing does not have pollen grain on the column cap, like Fig. 7; And a large amount of callose depositions are arranged on the column cap,, the no compatibility of this Chinese rose hybridization is described like Fig. 8.
Claims (6)
1. Chinese rose cross compatibility rapid identification method, the collection that it is characterized in that comprising pollen with preserve step, castrate step, artificial pollination step, sample to be identified cut step, it is characterized in that also comprising the following steps:
A, the sample to be identified that will cut are fixed in the FAA fixer, after 24 hours, transfer to mass concentration and are in 70% the ethanolic solution and preserve; Wherein, said FAA fixer is following proportioning:
Mass concentration is 37～40% formaldehyde 5～10 ml
Mass concentration is 70% ethanol 80～90 ml
Glacial acetic acid 5～10 ml;
B, sample that steps A was fixed by conventional water flushing after, place the NaOH solution of 8mol/L, 55～60 ℃ of water-baths 15～20 minutes were taken out, in 20～28 ℃ of held 3～3.5 hours;
C, the sample of step B was soaked 1 hour with running water at least, change running water therebetween 3～4 times, remove most of sodium hydroxide solution, transfer to mass concentration afterwards and be in 0.1% the aniline blue dye liquor and dyeed 2～3 hours;
D, get the dyeing sample of step C, by routine with fluorescence microscope column cap callose deposition situation, the pollen germination state, the pollen tube growth state, thereby identify the Chinese rose cross compatibility.
2. the method for claim 1, the glacial acetic acid that it is characterized in that said steps A is commercial analysis net product.
3. the method for claim 1 is characterized in that the collection of said pollen and preservation step are: select the plant of robust growth, select and treat open flower; And the mark of listing, maternal castration, bagging; To place 20～25 ℃ of ventilations to dry in the shade from the flower pesticide that the male parent flower is taken; Get loose powder, 4 ℃ of preservations, for use.
4. the method for claim 1; It is characterized in that said castration step is: select the flower that will open next day on the maternal plant to castrate, bagging afterwards, at this moment still incomplete development is ripe for stamen; The pollen that can not be scattered during castration, the operating time with afternoon 15:00～17:00 for well.
5. the method for claim 1 is characterized in that said artificial pollination step is: in 9:00-10:00 in the morning, take off the bagging in the maternal strain earlier; Dip in writing brush and to get the pollen that scatters in right amount; Smear pollination on maternal column cap, bagging kept 30～48 hours again.
6. the method for claim 1 is characterized in that the step that cuts of said sample to be identified is: take off bagging, the style after gynobase downcuts pollination is together with ovary.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100408990A CN102550396A (en) | 2012-02-22 | 2012-02-22 | Method for quickly identifying cross-compatibility of rose hybrida |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012100408990A CN102550396A (en) | 2012-02-22 | 2012-02-22 | Method for quickly identifying cross-compatibility of rose hybrida |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN102550396A true CN102550396A (en) | 2012-07-11 |
Family
ID=46398053
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN2012100408990A Pending CN102550396A (en) | 2012-02-22 | 2012-02-22 | Method for quickly identifying cross-compatibility of rose hybrida |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102550396A (en) |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568869A (en) * | 2014-12-12 | 2015-04-29 | 中国热带农业科学院橡胶研究所 | Prediction method for fertility rate of cross pollination of rubber tree |
| CN105409760A (en) * | 2015-12-04 | 2016-03-23 | 贵州省植物园 | Method for improving cross pollination success rate of Chinese rose |
| CN109258456A (en) * | 2018-11-06 | 2019-01-25 | 兰州市农业科技研究推广中心 | A kind of method of China's bitter water rose artificial pollination |
| CN110012748A (en) * | 2019-04-23 | 2019-07-16 | 南京林业大学 | It polymerize monogynaecial stripping means in gynoecium |
| CN110583634A (en) * | 2019-10-13 | 2019-12-20 | 安徽省农业科学院水稻研究所 | Preservation method for length of rice stigma |
| WO2021027866A1 (en) * | 2019-08-15 | 2021-02-18 | 中国农业大学 | Rna in-situ hybridization optimization method used for cucumber seedling gene mapping research |
| CN112903636A (en) * | 2021-01-15 | 2021-06-04 | 中国农业科学院蜜蜂研究所 | Method for rapidly determining pollination strength of crops |
| CN114088672A (en) * | 2021-11-12 | 2022-02-25 | 中国林业科学研究院林业研究所 | Method for quickly identifying compatibility of hybrid hazel pollination varieties in Pingyou |
| CN114766353A (en) * | 2022-05-09 | 2022-07-22 | 辽宁省经济林研究所 | Rapid screening method and application of male pollination plants in actinidia arguta orchard |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1821415A (en) * | 2006-03-07 | 2006-08-23 | 中国农业科学院油料作物研究所 | Rape transgenic method |
-
2012
- 2012-02-22 CN CN2012100408990A patent/CN102550396A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1821415A (en) * | 2006-03-07 | 2006-08-23 | 中国农业科学院油料作物研究所 | Rape transgenic method |
Non-Patent Citations (5)
| Title |
|---|
| 刘红明等: "6个月季品种的自交亲和性", 《湖南农业大学学报（自然科学版）》 * |
| 孙宪芝: "北林月季杂交育种技术体系初探", 《中国优秀硕士学位论文全文数据库（硕士） 农业科技辑》 * |
| 张颢等: "切花月季杂交结实性研究", 《西南农业大学学报（自然科学版）》 * |
| 申书兴: "《园艺植物育种学实验指导》", 31 August 2002, 中国农业大学出版社 * |
| 白锦荣: "部分蔷薇属种质资源亲缘关系分析及抗白粉病育种", 《中国博士学位论文全文数据库 农业科技辑》 * |
Cited By (12)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104568869A (en) * | 2014-12-12 | 2015-04-29 | 中国热带农业科学院橡胶研究所 | Prediction method for fertility rate of cross pollination of rubber tree |
| CN105409760A (en) * | 2015-12-04 | 2016-03-23 | 贵州省植物园 | Method for improving cross pollination success rate of Chinese rose |
| CN109258456A (en) * | 2018-11-06 | 2019-01-25 | 兰州市农业科技研究推广中心 | A kind of method of China's bitter water rose artificial pollination |
| CN110012748A (en) * | 2019-04-23 | 2019-07-16 | 南京林业大学 | It polymerize monogynaecial stripping means in gynoecium |
| CN110012748B (en) * | 2019-04-23 | 2021-10-01 | 南京林业大学 | Method for stripping single pistil in polymeric pistil |
| WO2021027866A1 (en) * | 2019-08-15 | 2021-02-18 | 中国农业大学 | Rna in-situ hybridization optimization method used for cucumber seedling gene mapping research |
| CN110583634A (en) * | 2019-10-13 | 2019-12-20 | 安徽省农业科学院水稻研究所 | Preservation method for length of rice stigma |
| CN110583634B (en) * | 2019-10-13 | 2022-01-14 | 安徽省农业科学院水稻研究所 | Preservation method of rice stigma |
| CN112903636A (en) * | 2021-01-15 | 2021-06-04 | 中国农业科学院蜜蜂研究所 | Method for rapidly determining pollination strength of crops |
| CN112903636B (en) * | 2021-01-15 | 2023-03-10 | 中国农业科学院蜜蜂研究所 | Method for rapidly determining pollination strength of crops |
| CN114088672A (en) * | 2021-11-12 | 2022-02-25 | 中国林业科学研究院林业研究所 | Method for quickly identifying compatibility of hybrid hazel pollination varieties in Pingyou |
| CN114766353A (en) * | 2022-05-09 | 2022-07-22 | 辽宁省经济林研究所 | Rapid screening method and application of male pollination plants in actinidia arguta orchard |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN102550396A (en) | Method for quickly identifying cross-compatibility of rose hybrida | |
| Cheng et al. | Interspecific hybrids between Dendranthema morifolium (Ramat.) Kitamura and D. nankingense (Nakai) Tzvel. achieved using ovary rescue and their cold tolerance characteristics | |
| CN101283669B (en) | Breeding technique of obtaining triploid grape and ploidy early identification using embryo | |
| Herrero et al. | Flowering, fruit set and development. | |
| Kenrick | Review of pollen–pistil interactions and their relevance to the reproductive biology of Acacia | |
| Tang et al. | Intergeneric hybridization and relationship of genera within the tribe Anthemideae Cass.(I. Dendranthema crassum (kitam.) kitam.× Crossostephium chinense (L.) Makino) | |
| He et al. | Pollen viability and stigma receptivity in Lilium during anthesis | |
| CN101243776A (en) | Cultivation method for cabbage type rape Isolated microspore plant strain | |
| Guo et al. | Inducing triploids and tetraploids with high temperatures in Populus sect. Tacamahaca | |
| ISHIZAKA et al. | Production of interspecific hybrids of Cyclamen persicum Mill. and C. hederifolium Aiton. by ovule culture | |
| Li et al. | Induction of unreduced megaspores in Eucommia ulmoides by high temperature treatment during megasporogenesis | |
| Tang et al. | Molecular cytogenetic identification and relationship of the artificial intergeneric hybrid between Dendranthema indica and Crossostephium chinense by GISH | |
| Light et al. | Factors affecting germinable seed yield in Cypripedium calceolus var. pubescens (Willd.) Correll and Epipactis helleborine (L.) Crantz (Orchidaceae) | |
| Sosnowska et al. | Application of in vitro pollination of opened ovaries to obtain Brassica oleracea L.× B. rapa L. hybrids | |
| CN102972296B (en) | Acacia rachii pollen in-vitro culture activity detection technology | |
| CN108901824A (en) | A kind of in-vitro verification method of grass family self-incompatibility phenotype | |
| CN109566399A (en) | A kind of method of S haplotype molecular marking supplementary breeding rape of self incompatible lin | |
| Souza et al. | In vitro pollination and fluorescence microscopy for characterization of gametophytic self-incompatibility of Coffea canephora Pierre ex A. Froehner | |
| He et al. | Haploid induction via in vitro gynogenesis in tomato (Solanum lycopersicum L.) | |
| CN109874671A (en) | A kind of abductive approach of pumpkin haplobiont and culture medium for the abductive approach | |
| Ren et al. | Efficient somatic embryogenesis and bulblet regeneration of the endangered bulbous flower Griffinia liboniana | |
| Dolezel et al. | A novel method to prepare slides for high resolution chromosome studies in Musa spp | |
| CN105104186B (en) | A kind of method obtaining Semen Sesami wild species and cultigen distant hybrid progeny using young embryo rescue techniques | |
| Chen et al. | Using giant scarlet runner bean (Phaseolus coccineus) embryos to dissect the early events in plant embryogenesis | |
| Giorgi et al. | Karyotype of globe artichoke (Cynara cardunculus var. scolymus): preliminary studies to define its chromosome morphology |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C12 | Rejection of a patent application after its publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20120711 |