CN112931193A - Field identification method for distinguishing Moli cytoplasm sterile plants and fertile plants of rape - Google Patents
Field identification method for distinguishing Moli cytoplasm sterile plants and fertile plants of rape Download PDFInfo
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- CN112931193A CN112931193A CN202110445324.6A CN202110445324A CN112931193A CN 112931193 A CN112931193 A CN 112931193A CN 202110445324 A CN202110445324 A CN 202110445324A CN 112931193 A CN112931193 A CN 112931193A
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- 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
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
- A01G22/15—Leaf crops, e.g. lettuce or spinach
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Abstract
The invention discloses a field identification method for distinguishing a sterile plant and a fertile plant of rape Moli cytoplasm1Then F is substituted1And performing second round hybridization with the sterile plant in the rape radish cytoplasm hybrid commodity rape, and judging the Moli cytoplasm sterile plant and fertile plant of the rape according to the fructification condition. The identification method avoids the problem that the judgment result of the material to be identified is not accurate only through the self-incompatibility due to the self-incompatibility gene.
Description
Technical Field
The invention belongs to the field of crossbreeding, and particularly relates to a field identification method for distinguishing a Muli cytoplasmic sterile line plant and a fertile plant of rape.
Background
Rape is a brassica plant in cruciferae and is one of the main oil crops in the world. The types of rape currently used in production are mainly cabbage type rape (Brassica napus L.), mustard type rape (Brassica juncea L.), and cabbage type rape (Brassica rapa L.). In the heterosis utilization of rapeCytoplasmic male sterility systems (CMS) are classified into class 2, one is sporophyte male sterility, such as Polima (Polima CMS), radish cytoplasm (Ogu CMS), etc., the male stamens of these sterile lines are degenerated completely, there is almost no visible pollen in the flowering phase, and the sterile and fertile plants can be identified by visual inspection. Another type of male sterility in oilseed rape is gametophytic male sterility, such as Molly (M. arvensis) CMS[1,2,3,4,5]The shape of the stamen of the sterile line is completely preserved, a great amount of pollen without activity can be seen in the stamen in the flowering phase, and the sterile plants and fertile plants can not be distinguished by visual inspection[4]. In the three-line breeding of rape, a maintainer line is used as a recurrent parent, and backcross is carried out for more than 5 generations with a sterile plant of a cytoplasmic male sterile system, and a backcross progeny is a sterile line corresponding to the maintainer line.
Sterile plants and fertile plants of the male sterility type of sporophytes such as rape bolima, radish cytoplasm and the like can be identified by observing and comparing the degradation degree of anthers and the existence of pollen in the flowering phase; however, the anthers of the sterile plants and fertile plants of the male sterile type of the rape gametophyte such as Muley CMS and the like have normal appearances and visible pollen, and the sterile plants and the fertile plants cannot be identified by visual inspection. The conventional identification methods comprise 2 methods: one is to take pollen to carry out acetic acid carmine dyeing in a laboratory, and to observe the coloring condition of pollen grains by a microscope for distinguishing; the second method is to perform bag-covering selfing and to identify the fruiting situation according to the selfing. The first method needs to be operated in a laboratory, certain experimental equipment and reagents are needed, and the identification threshold is high; the second method is simple, but when the breeding material has self-incompatibility genes, identification is carried out only through whether the selfing is fruitful, and certain misjudgment risk exists.
Reference documents:
1.Prakash S,Kirti P B,Bhat S R,et al.A Moricandia arvensis-based cytoplasmic male sterility and fertility restoration system in Brassica juncea[J].Theor Appl Genet,1998,97:488-492
2.Kirti P B,Prakash S,Gaikwad K,et al.Chloroplast substitution overcomes leaf chlorosis in a Moricandia arvensis-based cytoplasmic male sterile Brassica juncea[J].Theor Appl Genet,1998,97:1179-1182
3.Singh K H.Hybrid development in mustard(Brasica juncea L.)-A success story[J].Sarson News,2008,12(2):2-3
4.Bhat S R,Prakash S,Kirti P B,et al.Aunique introgression from Moricandia arvensis confers male fertility to two different cytoplasmic male sterile lines of Brassica juncea[J].Plant Breeding,2005,124:117-120
5. leishaolin, Ducaifu, Chenli, et al, research progress on the cytoplasmic male sterile system of Brassica juncea (Moricadia arvensis) [ J ] proceedings of oil crops in China, 2018,040(002): 297-301).
Disclosure of Invention
The technical problem to be solved by the invention is as follows: how to provide a method for identifying the Muli cytoplasmic sterile plant and the fertile plant of the rape by operating in the field.
The technical scheme of the invention is as follows: the field identification method for distinguishing the Muli cytoplasmic sterile plant and the fertile plant of the rape comprises the following steps:
first year
Sowing: respectively sowing the Muli cytoplasm hybrid rape and the pure line rape;
bagging: before the next year of bud emergence and flowering, bagging the main inflorescences of the pure rape variety plants by adopting a sulfuric acid paper bag;
artificial pollination: after blooming, taking flowers in the plant bags of the bagged rape pure line varieties, and performing artificial pollination by adopting an artificial emasculation hybridization mode and taking the rape pure line varieties as male parents as rape Moli cytoplasm hybrid plants;
uncovering the bag: after the rape is withered, uncovering a rape pure line variety and a sulfuric acid paper bag of the pollinated rape Moli cytoplasm hybrid plant in time;
harvesting: the hybrid seeds obtained on the female parent rape moly cytoplasm hybrid seed plants after maturation are F for the hybridization of the rape pure line varieties and the rape moly hybrid seed female parents1Seed generation;
second year
Sowing: f is to be1Respectively sowing the generation seeds and the rape radish cytoplasm hybrid commodity rapeseeds;
bagging: before the next year buds appear and bloom, bagging by adopting a sulfuric acid paper bag: selecting robust F1The generation plants are bagged for the main inflorescence, and the sterile plants are selected and bagged from rape radish cytoplasm hybrid commodity rape plants through visual inspection;
artificial pollination: after flowering, the cap bag F is taken out1Replacing plant flowers in the plant bag, adopting an artificial hybridization mode and using F1Artificial pollination is carried out on rape radish cytoplasm hybrid commercial rape sterile plants which take the generation plants as male parents as bags;
uncovering the bag: the rape withers and uncovers F in time1Sulfuric acid paper bags for the generation plants and the pollinated rape radish cytoplasm hybrid commercial rape sterile plants;
and (3) identification: visual inspection of bagging F1Covering the plant with the rape radish cytoplasm hybrid commodity rape seed bag inflorescence fructification after plant generation and pollination, F1The rape radish cytoplasmic hybrid commodity rape which is self-sterile and cannot bear pollen is also sterile Moli cytoplasmic; f1The rape radish cytoplasmic hybrid commodity rape plant which is normally selfed and fruited and simultaneously receives the pollen is also a moli cytoplasmic fertile plant.
Compared with the prior art, the invention has the following beneficial effects:
1. the sterile plants and fertile plants are identified by utilizing the characteristic that the sterile line non-viable pollen can not finish fertilization on flower stigmas of other rape plants.
2. The identification method avoids the problem that the judgment result of the material to be identified is not accurate only through the self-incompatibility due to the self-incompatibility gene.
3. The identification method adopts other sterile rape plants for auxiliary identification, does not adopt the rape with normal fertility as a material, saves the work of manually castrating the rape before the artificial pollination of the normal rape, and reduces the workload of identification.
Detailed Description
The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were all commercially available unless otherwise specified.
Preparing materials: rape moly cytoplasm hybrid seed; seeds of pure line rape; rape radish cytoplasm hybrid commercial rape seed. All three materials are commercially available. Commercial rapeseeds of Mori and radish cytoplasmic hybrids are abundant in India, and pure line varieties are available at home and abroad.
The identification method comprises the following steps:
first year
Sowing: sowing the Muli cytoplasm hybrid rape and the pure rape variety in a seedling transplanting mode or a direct seeding mode;
bagging: before the next year buds appear and blooms, the main inflorescences of the pure rape variety plants are bagged by adopting a sulfuric acid paper bag; preferably selecting a sulfuric acid paper bag with the width of 20-30cm and the length of 30-50 cm;
artificial pollination: after blooming, the flowers in the plant bags of the bagged rape pure line varieties are taken, and pollen is lightly smeared on the flower heads of the rape Moli cytoplasm hybrid plant by adopting an artificial emasculation hybridization mode and taking the rape pure line varieties as male parents to finish artificial pollination;
uncovering the bag: after the rape is withered, uncovering a rape pure line variety and a sulfuric acid paper bag of the pollinated rape Moli cytoplasm hybrid plant in time;
harvesting: when rape pure line variety is bagged for selfing and female parent hybridization siliques turn yellow, and seeds are black brown or brown, the seeds can be harvested; wherein the seeds harvested on the female parent are F hybridized with the female parent of the hybrid rape moral hybrid1Seed generation (hereinafter referred to as "F1Seed generation ");
second year
Sowing: f is to be1Respectively sowing the generation seeds and the rape radish cytoplasm hybrid commodity rapeseeds in a seedling transplanting mode or a direct seeding mode, wherein the sowing quantity is based on the guarantee that 200 healthy seedlings can be respectively obtained;
bagging: before the buds and flowers appear in the next year, the sulfuric acid paper bags are bagged, and the sizes of the paper bags are the same as those of the sulfuric acid paper bags; selecting robust F1The main inflorescences of the generation plants are bagged and listed by 20 plants, and the rape radish cytoplasm hybrid commodity is obtainedSelecting 20 sterile plants in the rapeseed plants by visual inspection, and bagging and listing the sterile plants;
artificial pollination: after flowering, the cap bag F is taken out1Replacing plant flowers in the plant bag, adopting an artificial hybridization mode and using F1Slightly smearing pollen on flower stigma of the bagged rape radish cytoplasmic hybrid commodity rape seed sterile plant by taking the generation plant as a male parent to finish artificial pollination;
uncovering the bag: after rape withers for 10 days, F is uncovered in time1Sulfuric acid paper bags for the generation plants and the pollinated rape radish cytoplasm hybrid commodity rape plants;
and (3) identification: visual inspection of bagging F1And (3) covering the plant generation and the pollinated rape radish cytoplasm hybrid commodity rape seed with a bag and bearing the seed sequence. F1The rape radish cytoplasmic hybrid commodity rapeseeds which are self-infertile and receive pollen are not fruitful and are Moli cytoplasmic sterile plants; f1The rape radish cytoplasmic hybrid commodity rape plants which are normally selfed and bear the pollen at the same time in the generation plants are also normally fruited as molli cytoplasmic fertile plants.
Claims (2)
1. The field identification method for distinguishing the Muli cytoplasmic sterile plant and the fertile plant of the rape is characterized by comprising the following steps of:
first year
Sowing: respectively sowing the Muli cytoplasm hybrid rape and the pure line rape;
bagging: before the next year of bud emergence and flowering, bagging the main inflorescences of the pure rape variety plants by adopting a sulfuric acid paper bag;
artificial pollination: after blooming, taking flowers in the plant bags of the bagged rape pure line varieties, and performing artificial pollination by adopting an artificial emasculation hybridization mode and taking the rape pure line varieties as male parents as rape Moli cytoplasm hybrid plants;
uncovering the bag: after the rape is withered, uncovering a rape pure line variety and a sulfuric acid paper bag of the pollinated rape Moli cytoplasm hybrid plant in time;
harvesting: the hybrid seeds harvested on the female parent rape moly cytoplasm hybrid plants after maturation are the hybrid seedsF for hybridization of rape pure line variety and rape moral hybrid1Seed generation;
second year
Sowing: f is to be1Respectively sowing the generation seeds and the rape radish cytoplasm hybrid commodity rapeseeds;
bagging: before the next year buds appear and bloom, bagging by adopting a sulfuric acid paper bag: selecting robust F1The generation plants are bagged for the main inflorescence, and the sterile plants are selected and bagged from rape radish cytoplasm hybrid commodity rape plants through visual inspection;
artificial pollination: after flowering, the cap bag F is taken out1Replacing plant flowers in the plant bag, adopting an artificial hybridization mode and using F1Artificial pollination is carried out on rape radish cytoplasm hybrid commercial rape sterile plants which take the generation plants as male parents as bags;
uncovering the bag: the rape withers and uncovers F in time1Sulfuric acid paper bags for the generation plants and the pollinated rape radish cytoplasm hybrid commodity rape plants;
and (3) identification: visual inspection of bagging F1Covering the plant with the rape radish cytoplasm hybrid commodity rape seed bag inflorescence fructification after plant generation and pollination, F1The rape radish cytoplasmic hybrid commodity rape sterile plant which is self-sterile and cannot bear the pollen is also non-sterile and is a moli cytoplasmic sterile plant; f1The rape radish cytoplasmic hybrid commodity rape sterile plant which is normally selfed and fruited and simultaneously receives the pollen in the generation plant also can fruitfully and normally becomes a moli cytoplasmic fertile plant.
2. The field identification method of claim 1, wherein in the second year, rape withers for 10 days, and the sulfuric acid paper bag is uncovered.
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102150613A (en) * | 2011-04-20 | 2011-08-17 | 西南大学 | Method for breeding cytoplasm male sterile line of novel cabbage type rape |
| CN102217523A (en) * | 2011-04-20 | 2011-10-19 | 华中农业大学 | Method for breeding rape temperature sensitive Polima cytoplasmic male sterile dual purpose line by microspore culturing process |
| CN102475058A (en) * | 2010-11-30 | 2012-05-30 | 华中农业大学 | Seed selection method of cabbage type brassica juncea cytoplasmic male sterile line |
| CN107896973A (en) * | 2017-11-16 | 2018-04-13 | 甘肃农业大学 | The selection of strong winter habit Chinese cabbage type winter rape thermosensitive type male sterility line |
| CN111109072A (en) * | 2020-01-02 | 2020-05-08 | 贵州省油料研究所(贵州省香料研究所) | Breeding method of purple-yellow leaf ornamental type rape line |
-
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- 2021-04-25 CN CN202110445324.6A patent/CN112931193A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102475058A (en) * | 2010-11-30 | 2012-05-30 | 华中农业大学 | Seed selection method of cabbage type brassica juncea cytoplasmic male sterile line |
| CN102150613A (en) * | 2011-04-20 | 2011-08-17 | 西南大学 | Method for breeding cytoplasm male sterile line of novel cabbage type rape |
| CN102217523A (en) * | 2011-04-20 | 2011-10-19 | 华中农业大学 | Method for breeding rape temperature sensitive Polima cytoplasmic male sterile dual purpose line by microspore culturing process |
| CN107896973A (en) * | 2017-11-16 | 2018-04-13 | 甘肃农业大学 | The selection of strong winter habit Chinese cabbage type winter rape thermosensitive type male sterility line |
| CN111109072A (en) * | 2020-01-02 | 2020-05-08 | 贵州省油料研究所(贵州省香料研究所) | Breeding method of purple-yellow leaf ornamental type rape line |
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| 巩振辉等: "《园艺植物育种学实验指导》", 31 August 2017, 西北农林科技大学出版社 * |
| 怎样种油菜编写组: "《怎样种油菜》", 31 January 1979, 上海人民出版社 * |
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