CN106258941A - The selection-breeding of a kind of Wuta-tsai and propagation method - Google Patents
The selection-breeding of a kind of Wuta-tsai and propagation method Download PDFInfo
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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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Abstract
The selection-breeding of a kind of Wuta-tsai and propagation method, the donor being cytoplasmic male sterile line sterile cytoplasm with cabbage cytoplasm male sterile line CMS65 7, purple leaf Chinese cabbage selfing line 08 131 and the donor that purple leaf mustard selfing line 08 550 is high anthocyanin character, the donor being Wuta-tsai character with Wuta-tsai 15 141, pass through distant hybridization, rescue culture, intervarietal hybridization, many for continuous backcross, in conjunction with experiment in cultivation, Disease Resistance Identification, anthocyanin content is identified and Fertility identification has obtained inheritance stability, disease resistance is strong, growth traits is excellent, Wuta-tsai cytoplasmic male sterile line and holding that anthocyanin content is high are.Have the advantages that anthocyanin content is high；Its fertility all shows 0 grade, and sterile plant rate and sterile degree are 100%.
Description
Technical field
The invention belongs to Cruciferae Vegetables in Brassica new variety selective breeding technology field, relate to a kind of Wuta-tsai selection-breeding and
Propagation method.
Background technology
Wuta-tsai (Brassica campestris L. ssp. chinensis var. rosularis Tsen, AA,
2n=20) it is a kind of Cruciferae Brassica genus pot-herb.The selection of Wuta-tsai kind generally has three kinds of approach now: one,
Conventional variety；Two, using cytoplasmic male sterile line is the maternal first generation of hybrid cultivated, and its sterile cytoplasm is mainly ogu CMS
With pol CMS；Three, the first generation of hybrid that self incompatible line is cultivated is utilized for female parent.Wherein, with conventional variety and cytoplasmatic male
The first generation of hybrid of sterile line breeding accounts for the major part of Wuta-tsai kind.Utilize the F of cytoplasmic male sterile line selection-breeding1Because of excellent
Hybrid vigor, in terms of the resistance and yield of kind, all have good performance.In the new varieties choosing taken as the leading factor with male sterility
In educating, long-term single kytoplasm type easily causes sickness rate to be increased, and then affects crop yield；Ogu CMS kytoplasm honey simultaneously
Gland is degenerated, and causes hybrid seed yield low, and pol CMS kytoplasm poor stability, fertility is easily subject to the impact of temperature, reduces hybrid pure
Degree.Therefore find in cytoplasmic male sterile line for maternal heterosis utilization and use more cytoplasmatic male not
Educating type is one the long-term job content of breeding man.
Anthocyanin is a kind of important Secondary metabolites, and its main component is flavonoid.According to research for many years
As a result, anthocyanin has health care biology, such as antitumor, anticancer, delaying senility, the liver protecting in the diet of the mankind
Deng.Thus, cultivate vegetable and the cereal crops of high anthocyanin content for many years, become plant variety selection-breeding breeding objective it
One.In the vegetable variety selection-breeding of high anthocyanin content, in view of anthocyanin is present in the blade of plant more, its phenotype is blade
Present purple, red or blue, thus the kind that selection-breeding anthocyanin content is high in leaf vegetables has the advantage of uniqueness.
In brassica plant, according to carrying out intervarietal hybridization between the theoretical multiple species of Yu Shi triangle, thus be easy to by multiple not
Gather in a kind with the anthocyanin expressing gene in source.Wuta-tsai is as a kind of main Vegetables in Brassica, and selection-breeding is high
The sterile line of anthocyanin content is significant for selection-breeding height anthocyanin content kind.
Summary of the invention
In order to overcome the disadvantages mentioned above of prior art, a kind of new sterility of selection-breeding of the present invention is stable, thoroughly, not by temperature
And the selection-breeding of the Wuta-tsai of lighting conditions and propagation method, this sterile line has that anthocyanin content is high, anthocyanin contains simultaneously
The features such as amount is stable, for cultivating the germ plasm resource that the Wuta-tsai new varieties of high anthocyanin content provide new.
The technical solution adopted for the present invention to solve the technical problems is: the selection-breeding of a kind of Wuta-tsai, with Chinese cabbage cell
Matter male sterility line CMS65-7 be the donor of cytoplasmic male sterile line sterile cytoplasm, purple leaf (high anthocyanin content) Chinese cabbage from
Friendship be 08-131 and purple leaf (high anthocyanin content) Caulis et Folium Brassicae junceae selfing line 08-550 be the donor of high anthocyanin character, with Wuta-tsai
15-141 is the donor of Wuta-tsai character.By distant hybridization, rescue culture, intervarietal hybridization, how for continuous backcross, combine cultivation and try
Test, Disease Resistance Identification, anthocyanin content identify and Fertility identification has obtained inheritance stability, disease resistance is strong, growth traits is excellent, flower
The high Wuta-tsai cytoplasmic male sterile line of color glycosides content and keep system, this sterile line be different from Ogu CMS, nap CMS,
The Wuta-tsai cytoplasmic male sterile line that the novel anthocyanin content of tour CMS is high.
A kind of propagation method of Wuta-tsai, concrete propagation steps is as follows:
1, with cabbage cytoplasm male sterile line CMS65-7 as female parent, with purple leaf Chinese cabbage selfing line 08-131 as male parent, by two
In person filial generation, the sterile individual plant of high anthocyanin is as nonrecurrent parent, backcrosses for 8 generations with Wuta-tsai 15-141 for recurrent parent,
I.e. [(CMS65-7 × 08-131) × 15-141] BC8F1, complete 08-131 height anthocyanin character, Wuta-tsai 15-141 and CMS65-
The gene pyramiding of 7 sterile cytoplasms, the numbered CMS10-122 of offspring；
2, with purple leaf mustard 08-550 as female parent, carry out rescue culture after carrying out distant hybridization with Wuta-tsai 15-141, obtain F1；With
F1Individual plant is nonrecurrent parent, and 15-141 is that recurrent parent backcrosses after 8 generations, select purple leaf individual plant selfing again obtain (08-550 ×
15-141) BC8F2, plant (08-550 × 15-141) BC8F2, take offspring and do not occur the parent of purple leaf trait segregation to be future
Male parent, completes the Wuta-tsai strain breeding containing 08-550 height anthocyanin gene, numbered 15-788；
3, with CMS10-122 as female parent, 15-788 is male parent, the high anthocyanin that the seed on results female parent is in the present invention
The Wuta-tsai sterile line of content, numbered CMS15-33, the seed on results 15-788 is the holding system of the present invention.
The positive effect of the present invention is:
(1) the Wuta-tsai cytoplasmic male sterile line of the present invention has been polymerized the different high flower deriving from purple leaf mustard and purple leaf Chinese cabbage
Color glycosides expressing gene, has the advantages that anthocyanin content is high；
(2) sterility of the Wuta-tsai cytoplasmic male sterile line of the present invention stable and thoroughly, not by temperature and illumination condition shadow
Ringing, the Fertility identification in China's Shaoguan (short-day) and Gansu and political affairs (long-day) 8 generations plantation in continuous 4 years shows, its fertility is equal
Showing 0 grade, sterile plant rate and sterile degree are 100%.
Accompanying drawing explanation
Fig. 1 is present invention process flow chart.
Detailed description of the invention
See Fig. 1, the selection-breeding of a kind of Wuta-tsai, first determine purple leaf Chinese cabbage selfing with customary physiological biochemical analysis method
It is 08-131 and the anthocyanin content of purple leaf mustard selfing line 08-550 and constituent thereof, then with classical genetics analysis side
Method determines the genetic development of regulation and control 08-131 and 08-550 purple leaf character gene, so that it is determined that control 08-131 and 08-550
The gene of purple leaf character be 1 pair of Gene Handling complete dominance character and both be non-homogeneous gene；Male with cabbage cytoplasm
Sterile line CMS65-7 is the donor of cytoplasmic male sterile line sterile cytoplasm, purple leaf (high anthocyanin content) Chinese cabbage selfing line 08-
131 and purple leaf (high anthocyanin content) Caulis et Folium Brassicae junceae selfing line 08-550 are the donor of high anthocyanin character, with Wuta-tsai 15-141 are
The donor of Wuta-tsai character.By distant hybridization, rescue culture, intervarietal hybridization, many for continuous backcross, in conjunction with experiment in cultivation, resistance
Identify, anthocyanin content identifies and Fertility identification has obtained inheritance stability, disease resistance is strong, growth traits is excellent, anthocyanin content
High Wuta-tsai cytoplasmic male sterile line and keep system, this sterile line is different from Ogu CMS, nap CMS, tour CMS
The Wuta-tsai cytoplasmic male sterile line that novel anthocyanin content is high.
The propagation method of a kind of Wuta-tsai, specifically comprises the following steps that
Early stage physiological and biochemical analysis:
One, purple leaf mustard and purple leaf Chinese cabbage anthocyanin assay
The purple leaf mustard selfing line 08-550 of plantation and purple leaf Chinese cabbage selfing line 08-131 respectively, in 5 leaf phase purple leaf trait expression
For time obvious, take big SANYE, use spectrophotography to measure its anthocyanin content respectively.
Two, purple leaf mustard and purple leaf Chinese cabbage anthocyanin compound mensuration
1, anthocyanin in purple leaf mustard selfing line 08-550 and purple leaf Chinese cabbage selfing line 08-131 is extracted respectively at 5 leaf phases；
2, LC-MS method is used to carry out component analysis the anthocyanin extracted in step 1；
Determine that purple leaf mustard 08-550 and purple leaf Chinese cabbage 08-131 is mainly composed of anthocyanin according to result in, two, simultaneously its
Constituent is different, so that it is determined that the gene controlling purple leaf mustard 08-550 and purple leaf Chinese cabbage 08-131 anthocyanin synthesis is different
Gene.
Three, the genetic analysis of 08-550 purple leaf character
1, with purple leaf mustard 08-550 and greenery Caulis et Folium Brassicae junceae 08-332 as parent, carry out reciprocal crosses respectively, obtain F1；
2, F in plantation 11, find the purple that leaf color is all identical with 08-550, separate without leaf color；
3, by F1Selfing, respectively obtains 3 F2Colony, plants F2Colony, finds that offspring's purple leaf and greenery segregation ratio are 3:1；
4, with F1For female parent, 08-332 is male parent, uses the artificial method dialling flower bud producing Crossbred to obtain BC1F1Individual plant, BC1F1No
It is purple leaf with strain leaf color Phenotype: greenery=1:1；
5, above-mentioned result of the test shows, the complete dominance character that purple leaf character is 1 pair of Gene Handling of 08-550.
Four, the genetic analysis of 08-131 purple leaf character
1, with purple leaf Chinese cabbage 08-131 and greenery Chinese cabbage 08-426 as parent, carry out reciprocal crosses respectively, obtain F1；
2, F in plantation 11, find that leaf color is the purple identical with 08-131, separate without leaf color；
3, by F1Selfing, respectively obtains 3 F2Colony, plants F2Colony, finds that offspring's purple leaf and greenery segregation ratio meet 3:1 and lose
Pass rule；
4, with F1For female parent, 08-426 is male parent, uses the artificial method dialling flower bud producing Crossbred to obtain BC1F1Individual plant, BC1F1No
It is purple leaf with strain leaf color Phenotype: greenery=1:1；
5, the above results shows, 08-131 purple leaf character is the complete dominance character of 1 pair of Gene Handling.
Sterile line and holding system breeding concrete grammar:
One, the selection-breeding of 15-141
1, the Wuta-tsai selfing line 10-222 of our unit's selection-breeding is hybridized with 10-327 emasculation, obtain F1；
2, by F1Selfing obtains F2, at F2Carrying out disease resistance test in colony and plant type is observed, disease resistance is good, plant type is excellent in selection
Multiple individual plants as F3Enter next step selection-breeding；
3, at F3In colony, carry out disease resistance test further and plant type is observed, select excellent F4Individual plant carries out next step selection-breeding；
4, at F4Carrying out disease resistance and plant type viewing test in individual plant, carry out yielding ability test simultaneously, selection disease resistance is good, plant type
And the F that yield is all good5Individual plant, numbered 15-141.
Two, holding is the selection-breeding of 15-788
1, with purple leaf mustard selfing line 08-550 as female parent, with 15-141 as male parent, carry out embryo after using artificial emasculation hybridization and draw
Rescue, it is thus achieved that (08-550 × 15-141) F1；
2, with (08-550 × 15-141) F1For female parent, with 15-141 as male parent, the method for artificial emasculation hybridization is used to carry out back
Hand over, it is thus achieved that [(08-550 × 15-141) × 15-141] BC1F1Individual plant；
3, [(08-550 × 15-141) × 15-141] BC that will obtain in 21F1[(08-550 × 15-141) is obtained after selfing
×15-141]BC1F2, at [(08-550 × 15-141) × 15-141] BC1F2The individual plant that in colony, selection purple is the deepest is as mother
This, with 15-141 as male parent, use the method for artificial emasculation hybridization to obtain filial generation [(08-550 × 15-141) × 15-
141]BC2F1；
4, with [(08-550 × 15-141) × 15-141] BC in 32F1The purple the deepest individual plant of leaf is maternal, and 15-141 is male parent,
The method using artificial emasculation hybridization backcrosses, it is thus achieved that [(08-550 × 15-141) × 15-141] BC3F1；
5, with [(08-550 × 15-141) × 15-141] BC in 43F1The deepest purple individual plant of phenotype is maternal, and 15-141 is father
This, use artificial emasculation method to backcross, it is thus achieved that [(08-550 × 15-141) × 15-141] BC4F1；
6, as figure 5 illustrates, in filial generation, the deepest purple individual plant of color is shown as nonrecurrent parent, with 15-141 as samsara
Parent, backcrosses 8 times, it is thus achieved that [(08-550 × 15-141) × 15-141] BC8F1；
7, by [(08-550 × 15-141) × 15-141] BC8F1Colony is in units of individual plant, and mark is carried out in bagging selfing simultaneously,
Obtain [(08-550 × 15-141) × 15-141] BC8F2Strain, by [(08-550 × 15-141) × 15-141] BC8F2Each
Taking the plantation of Some seeds duplicate rows in strain, selecting phenotype is purple blade, does not has the strain of trait segregation simultaneously, determines this strain
For the individual plant that isozygotys, using this individual plant as holding it is, numbered 15-788.
Three, the selection in sterile cytoplasm source
The cabbage cytoplasm male sterile line (patent publication No. is: CN 1849881A) of selection Hua Zhong Agriculture University selection-breeding is
Female parent material, through Fertility identification and resistant proof, growing way observation, it is female for selecting the sterile individual plant that growing way is excellent, disease resistance is good
This numbered CMS63-5.
Four, the selection-breeding of purple leaf Wuta-tsai sterile line CMS10-122
1, with CMS63-5 as female parent, purple leaf Chinese cabbage selfing line 08-131 is male parent, uses the artificial method dialling flower bud pollination to obtain miscellaneous
Hand over offspring (CMS63-5 × 08-131) F1；
2, to (CMS63-5 × 08-131) F1Carrying out Fertility identification and Phenotypic Observation, select sterility good, purple leaf character is the most obvious
Individual plant be maternal, with 15-141 as male parent, use the artificial method dialling flower bud pollination to obtain [(CMS63-5 × 08-131) × 15-
141]F1, 15-141 bagging selfing simultaneously is reserved seed for planting standby；
3, by [(CMS63-5 × 08-131) × 15-141] F1And the community plantation of 15-141 colony, carry out leaf color observation, disease resistance
Test, plant type are observed and Fertility identification, select purple leaf character the most substantially, plant type is good, disease resistance by force, the most sterile individual plant be mother
This, select disease resistance in 15-141 good, and the individual plant that plant type is excellent is male parent, uses group flower bud artificial pollination mode to hybridize, obtains
Obtain [(CMS63-5 × 08-131) × 15-141] BC1F1Colony；
4, step in 3 is repeated, with [(CMS63-5 × 08-131) × 15-141] BC1F1The middle purple the most obvious sterile individual plant of leaf is
Female parent, 15-141 is male parent, it is thus achieved that [(CMS63-5 × 08-131) × 15-141] BC2F1Colony；
5, step in 4 is repeated, with [(CMS63-5 × 08-131) × 15-141] BC2F1The middle purple the most obvious sterile individual plant of leaf is
Female parent, 15-141 is male parent, it is thus achieved that [(CMS63-5 × 08-131) × 15-141] BC3F1Colony；
6, on the basis of 8 generations that backcrossed, it is thus achieved that [(CMS63-5 × 08-131) × 15-141] BC8F1Sterile population, it is numbered
CMS10-122。
Five, Wuta-tsai sterile line CMS15-33 and the expanding propagation of holding system
1, CMS10-122 and 15-788 interlacing being planted, column number proportion is 3:1, treats that initial bloom stage carries out breeding with guard method；
2, respectively results CMS10-122 and 15-788 seed, Second Year carry out respectively anthocyanin content qualification, Disease Resistance Identification,
Growing way measure and Fertility identification, all sterile individual plants of seed that result CMS10-122 is gathered in the crops, simultaneously blade dark purple,
Anthocyanin content is high, and the 15-788 upper results all individual plants of educating of seed, blade presents purple and growing way is good, and disease resistance is strong.
Sterile line during gathered in the crops seed is the present invention on CMS10-122, numbered CMS15-33, on 15-788, gathered in the crops seed is this
Holding system in invention.
Claims (2)
1. a selection-breeding for Wuta-tsai, is characterized in that: with cabbage cytoplasm male sterile line CMS65-7 for cytoplasmatic male not
Educate is the donor of sterile cytoplasm, high anthocyanin content purple leaf Chinese cabbage selfing line 08-131 and high anthocyanin content purple leaf mustard selfing
Be 08-550 be high anthocyanin character donor, be Wuta-tsai character with Wuta-tsai 15-141 donor, by distant hybridization, embryo
Redemption, intervarietal hybridization, how for continuous backcross, in conjunction with experiment in cultivation, Disease Resistance Identification, anthocyanin content identifies and Fertility identification obtains
Arrived inheritance stability, disease resistance is strong, growth traits is excellent, anthocyanin content is high Wuta-tsai cytoplasmic male sterile line and holding
System, this sterile line is different from the high Wuta-tsai Cytoplasm of novel anthocyanin content of Ogu CMS, nap CMS, tour CMS
Male sterility line.
The propagation method of a kind of Wuta-tsai the most as claimed in claim 1, it is characterised in that concrete propagation steps is as follows:
1, with cabbage cytoplasm male sterile line CMS65-7 as female parent, with purple leaf Chinese cabbage selfing line 08-131 as male parent, by two
In person filial generation, the sterile individual plant of high anthocyanin is as nonrecurrent parent, backcrosses for 8 generations with Wuta-tsai 15-141 for recurrent parent,
I.e. [(CMS65-7 × 08-131) × 15-141] BC8F1, complete 08-131 height anthocyanin character, Wuta-tsai 15-141 and CMS65-
The gene pyramiding of 7 sterile cytoplasms, the numbered CMS10-122 of offspring；
2, with purple leaf mustard 08-550 as female parent, carry out rescue culture after carrying out distant hybridization with Wuta-tsai 15-141, obtain F1；With
F1Individual plant is nonrecurrent parent, and 15-141 is that recurrent parent backcrosses after 8 generations, select purple leaf individual plant selfing again obtain (08-550 ×
15-141) BC8F2, plant (08-550 × 15-141) BC8F2, take offspring and do not occur the parent of purple leaf trait segregation to be future
Male parent, completes the Wuta-tsai strain breeding containing 08-550 height anthocyanin gene, numbered 15-788；
3, with CMS10-122 as female parent, 15-788 is male parent, the high anthocyanin that the seed on results female parent is in the present invention
The Wuta-tsai sterile line of content, numbered CMS15-33, the seed on results 15-788 is the holding system of the present invention.
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Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1849881A (en) * | 2006-06-05 | 2006-10-25 | 华中农业大学 | Selective breeding method for cabbage cytoplasm male sterile line |
| WO2011031680A1 (en) * | 2009-09-11 | 2011-03-17 | Mendel Biotechnology, Inc. | Enhancement of plant yield, vigor and stress tolerance ii |
| CN103444515A (en) * | 2013-08-26 | 2013-12-18 | 安徽省农业科学院园艺研究所 | Transfer method for male sterile line of purple wuta-tsai |
| CN103461100A (en) * | 2013-08-26 | 2013-12-25 | 安徽省农业科学院园艺研究所 | Purple Wuta-tsai breeding method |
| WO2014195152A1 (en) * | 2013-06-04 | 2014-12-11 | Bayer Cropscience Nv | Improved hybrid seed production method |
| CN104521740A (en) * | 2014-12-17 | 2015-04-22 | 安徽农业大学 | Synchronous transform-breeding method for wuta-cai homozygous inbred line and cytoplasmic male sterile line |
| WO2016067284A1 (en) * | 2014-10-28 | 2016-05-06 | Equi-Nom Ltd. | Production of hybrid seeds lot using natural pollination |
| CN105815211A (en) * | 2016-03-26 | 2016-08-03 | 安徽科技学院 | Method for improving self-incompatibility Wuta-tsai hybridization efficiency |
-
2016
- 2016-09-21 CN CN201610837233.6A patent/CN106258941A/en active Pending
Patent Citations (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1849881A (en) * | 2006-06-05 | 2006-10-25 | 华中农业大学 | Selective breeding method for cabbage cytoplasm male sterile line |
| WO2011031680A1 (en) * | 2009-09-11 | 2011-03-17 | Mendel Biotechnology, Inc. | Enhancement of plant yield, vigor and stress tolerance ii |
| WO2014195152A1 (en) * | 2013-06-04 | 2014-12-11 | Bayer Cropscience Nv | Improved hybrid seed production method |
| CN103444515A (en) * | 2013-08-26 | 2013-12-18 | 安徽省农业科学院园艺研究所 | Transfer method for male sterile line of purple wuta-tsai |
| CN103461100A (en) * | 2013-08-26 | 2013-12-25 | 安徽省农业科学院园艺研究所 | Purple Wuta-tsai breeding method |
| WO2016067284A1 (en) * | 2014-10-28 | 2016-05-06 | Equi-Nom Ltd. | Production of hybrid seeds lot using natural pollination |
| CN104521740A (en) * | 2014-12-17 | 2015-04-22 | 安徽农业大学 | Synchronous transform-breeding method for wuta-cai homozygous inbred line and cytoplasmic male sterile line |
| CN105815211A (en) * | 2016-03-26 | 2016-08-03 | 安徽科技学院 | Method for improving self-incompatibility Wuta-tsai hybridization efficiency |
Non-Patent Citations (3)
| Title |
|---|
| HAI‑LONG YU: "Development of a novel allele‑specific Rfo marker and creation of Ogura CMS fertility‑restored interspecific hybrids in Brassica oleracea", 《THEOR APPL GENET》 * |
| 宋波等: "乌塌菜Ogura雄性不育系组合与保持系组合杂种优势比较", 《江苏农业科学》 * |
| 宋波等: "我国乌塌菜研究进展", 《中国蔬菜》 * |
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