CN106171948A - A kind of selection of wide handle mustard cytoplasm male sterility line - Google Patents
A kind of selection of wide handle mustard cytoplasm male sterility line Download PDFInfo
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- CN106171948A CN106171948A CN201610542024.9A CN201610542024A CN106171948A CN 106171948 A CN106171948 A CN 106171948A CN 201610542024 A CN201610542024 A CN 201610542024A CN 106171948 A CN106171948 A CN 106171948A
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- mustard
- wide handle
- male sterility
- sterility line
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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
- 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
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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- Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses the selection of a kind of wide handle mustard cytoplasm male sterility line, comprise the steps: to choose tumorous stem mustard (hot pickled mustard tube) cytoplasm male sterility line JL154 5A as female parent, and choose wide handle mustard Elite inbred and carry out conventional hybridization as transformation male parent and obtain F1, offspring selects typical case's individual plant and male parent continuous backcross 7 generation, obtaining sterile character stable, wide handle mustard cytoplasm male sterility line and holding that economic characters are good are.The present invention is with tumorous stem mustard cytoplasm male sterility line as sterile source, wide handle mustard elite germplasm is male parent, by hybridization and continuous backcross, at home and abroad taking the lead in cultivating wide handle mustard cytoplasm male sterility line and the holding system that sterile character is stablized, economic characters are good and differs from one another, the cross-breeding genetic germplasm and the genus that enrich mustard vegetables (wide handle mustard) are initiated both at home and abroad；Realize this crop " three are " supporting, and then cultivate the wide handle mustard miscellaneous generation new varieties that anti-(resistance to) characteristic of disease of high yield and high quality is stronger, it is achieved heterosis utilization, there is the market prospect extensively read.
Description
Technical field
The present invention relates to Cruciferae Brassica genus mustard vegetables breeding technique, be specifically related to a kind of wide handle mustard cytoplasmic male
Sterile line breeding method.
Background technology
Wide handle mustard (Brassica juncea var.latipa), is Cruciferae Brassica genus Caulis et Folium Brassicae junceae kind leaf mustard class vegetable
One mutation, is commonly called as bamboo shell dish, green vegetable, wide plate dish, because its petiole and the roomy plumpness of middle rib, quality are tender and crisp, simultaneously containing different journeys
The acid of degree, boil soup or mutton cooked in a chafing pot taste is the most beautiful, have and reduce phlegm and internal heat, imperial rheumatism, profit the five internal organs, mend the work of sky sun blood circulation promoting
Effect, is quite liked by common people, is that " primary raw material is also that the characteristic that citizen commonly use eats vegetable raw to well-known Sichuan acid (bubble) dish.
Extensively cultivating in urban district, south China each province at present, Annual planting area reaches more than 1,500,000 mu, especially with faces such as Sichuan, Hunan, Guizhou
Long-pending relatively big, produce plantation processing and concentrate the most.
At present, have not been reported with research about wide handle mustard cytoplasm male sterility line selection-breeding both at home and abroad.The widest handle mustard
Large area produces by kind of an aspect, and for a long time, either province or the Chongqing City such as river, Hunan, Guizhou Province, be all directly to use performance the best
Local varieties, how mixed and disorderly " " and " inbreeding depression " phenomenon that large area produces be very universal, and direct result is exactly that raw material is made
The growth of thing is irregular, yield is the highest, poor quality, disease-resistant (inverse) property are weak, seriously governs the rapid and healthy of this industry.
Summary of the invention
For solving the problems referred to above, the invention provides the selection of a kind of wide handle mustard cytoplasm male sterility line, utilize mustard
Dish class vegetable (tumorous stem mustard) sterile line is sterile source, selects typical case's individual plant by hybridization, offspring and divides with male parent continuous backcross and SSR
Sub-Marker Identification, has cultivated the wide handle mustard cytoplasm male sterility line that sterile character is stable, economic characters are good and differs from one another
And keeping system, it is achieved this crop " three are " is supporting, and then cultivates the wide handle mustard hybridization new product that anti-(resistance to) characteristic of disease of high yield and high quality is strong
Kind, it is achieved heterosis utilization, belong to the most pioneering.
For achieving the above object, the technical scheme that the present invention takes is:
The selection of a kind of wide handle mustard cytoplasm male sterility line, comprises the steps:
Choose tumorous stem mustard cytoplasm male sterility line JL154-5A as female parent, and choose wide handle mustard Elite inbred simultaneously and make
Carrying out conventional hybridization for transformation male parent and obtain F1, offspring selects typical case's individual plant and male parent continuous backcross 7 generation, it is thus achieved that sterile character is steady
Fixed, wide handle mustard cytoplasm male sterility line and holding that economic characters are good are.
The method have the advantages that
1, with tumorous stem mustard cytoplasm male sterility line as sterile source, wide handle mustard elite germplasm is male parent, by hybridization with continuous
Backcross, at home and abroad take the lead in cultivating sterile character is stable, economic characters are good and differs from one another wide handle mustard cytoplasmic male not
Educate and be and keep system, enrich the cross-breeding genetic germplasm of mustard vegetables and belong to the most pioneering.
2, based on the wide handle mustard cytoplasm male sterility line cultivated and holding system, it is achieved this crop " three are " is supporting,
And then cultivate the wide handle mustard miscellaneous generation new varieties that anti-(resistance to) characteristic of disease of high yield and high quality is strong, it is achieved and heterosis utilization, have wide
Market prospect.
Accompanying drawing explanation
Fig. 1 is the family tree of the selection of a kind of wide handle mustard cytoplasm male sterility line of the embodiment of the present invention.
Fig. 2 is that embodiment of the present invention primer " KBJ416 " at wide handle mustard sterile line and keeps polymorphism scattergram in system.
Detailed description of the invention
In order to make objects and advantages of the present invention clearer, below in conjunction with implementing example, the present invention is entered one
Step describes in detail.Should be appreciated that and described herein be embodied as example only in order to explain the present invention, be not used to limit this
Invention.
As it is shown in figure 1, embodiments provide the selection of a kind of wide handle mustard cytoplasm male sterility line, including such as
Lower step:
S1, with Chongqing southeast academy of agricultural sciences model tumorous stem mustard (hot pickled mustard tube) cytoplasm male sterility line cultivated such as the reddest
" JL154-5A " makees female parent, in March, 2011, at field system identification and confirm the basis that " JL154-5A " sterile character is stable
On, select various trait, different ecological type, wide handle mustard Elite inbred DJ, K6, HK-1, HK-2, K46, BK of different ripe property
Carrying out conventional hybridization as male parent and obtain F1, offspring selects typical case's individual plant and male parent continuous backcross 7 generation, it is thus achieved that sterile character is stable,
Wide handle mustard cytoplasm male sterility line that economic characters are good and keep system: DJA and DJB, K6A and K6B, HK-1A and HK-1B,
K46A and K46B, HK-2A and HK-2B, BKA and BKB；
In October, 2010, by tumorous stem mustard sterile line " JL154-5A ", is experimental field sowed in Chongqing southeast academy of agricultural sciences
Culturing and transplanting seedlings, has carried out observing qualification, really in florescence to the floral organ structure of sterile line, sterile trait expression in March, 2011
After card is the sterile line that sterility is thorough, character is stable, makees female parent with tumorous stem mustard sterile line JL154-5A, choose 6 excellent width
Handle mustard selfing line, every selfing line 3 individual plants, the most respectively paired cross, obtain F1 generation.Summer in August, 2011 is numerous, selects from F1 generation
Selecting 3 typical individual plants and be a mother this, backcross with the selfing line width handle mustard S1 of above-mentioned correspondence, the most maternal individual plant selfing measures its knot
Reality.The spring of continuous 4 years (2011-2014) and summer, the wide handle mustard selfing line of continuation is made samsara male parent and is backcrossed therewith, simultaneously
Maternal individual plant bagging selfing, measures its fertility.To in May, 2015 selected sterile degree, sterile plant rate is 100%, and economical
Wide handle mustard cytoplasm male sterility line and holding that character is the most stable are 6
S2, at hybridization and each backcross generations, the florescence, in units of material, carries out individual plant bagging selfing, examination sterile plant from
The fecundity of flower pollination.
S3, to backcross 7 generations (sterile line) and male parent thereof (keeping system), the typical individual plant of random selection 3-5 divides at full-bloom stage
Other Observe and measure corolla, stamen, gynoecium and the form and dimension of nectary, and its Flowering Characteristics is carried out systematic observation.
S4, pollen to backcross 7 generations (sterile line) and male parent thereof (keeping system), red tetrazolium (TTC) liquid with 0.5% is 38
Constant temperature dyeing 50min (dividing) at DEG C, measure its vitality.
S5, to sterile line and Bao Te system, carry out positive season plantation, conventional pipe at Chongqing southeast academy of agricultural sciences (height above sea level 273m)
Reason, when each material is buddingged, 5 typical individual plants of every material selection, by " mustard vegetables field test observes record project and mark
Accurate ", its morphological characteristic of systematic observation and major traits performance.
2014 days JIUYUE～in February, 2015, to being complete backcross transformation and reaching the institute of sterile line standard through Fertility identification
There is sterile line and keep system, concentrating on and cultivate, to its morphological characteristic and Main Agronomic under the same ecological condition in southeast academy of agricultural sciences, Chongqing
Character has carried out the observation of system and has identified.Result is:
The sterile strain of wide handle after continuous 7 generation backcross transformations, it is at leaf, leaf margin, middle arteries and veins color, blade spiny hair wax
The aspects such as powder, individual plant effective blade number, same or like with corresponding holding system performance.But it is fresh at plant height, degree of development, individual plant
The aspects such as weight, middle rib width and thickness, have bigger difference with the corresponding system that keeps.That is: sterile line keeps the plant of system than correspondence
High 2-4cm, the big 4-9cm of degree of development, single-strain fresh weight relatively keeps being weight 100-300g etc..Meanwhile, ratio vegetative growth phase of sterile line
Keep system long 3-5 days.
The 3-5 month in 2014, while proceeding wide handle mustard sterile line backcross transformation, to sterile line and keep accordingly be
Flower organ morphology feature, Flowering Characteristics and the florescence change carried out the observation of system.
(1) the floral organ feature of sterile line
The corolla keeping system is made up of 4 petals, and in cross, yellow or light yellow, 6 pieces of stamen, semi-finals 2 are weak；Gynoecium list
Raw, the many mastoid processes of stigma surface, the length of flower pesticide and filigree is slightly larger than the length of style, and flower pesticide is fuller；Nectary development is good, 3-
4, circular, green or light green color, indivedual bottle greens are spent and corresponding holding through the wide handle mustard sterile line of continuous backcross 7 generation transformation
System compares, and has the difference in its morphological characteristic and structure: when sterile line corolla contains flower, petal shows as half-open or standard-sized sheet, in not
The crosswise of rule；5-6 piece of every colored stamen, but flower pesticide is shrivelled, fine, have rib or many ribs；Filigree is short 0.87-than holding
1.45mm；Gynoecium Dan Sheng, the many mastoid processes of stigma surface, ovary mode of appearance and size are essentially identical with holding；Nectary development is good,
Notable difference is not had with corresponding holding system.
(2) Flowering Characteristics of sterile line
Through the wide handle mustard sterile line of continuous 7 generation backcross transformations, the almost stigma of every flower, just stretched out before alabastrum maturation
Reach 2.1-3.1mm outside flower bud, form special " the heterostyle phase ", the most manually award and can normally tie with normal mustard pollen
Setting seeds in fact, average setting percentage reaches more than 90%.Stigma appearing 2～3d (my god) after, petal the most entirely launches or half launches, in irregularly
Cross, then through 2～3d (my god) petal comes off, stigma surface blackening.And the holding of correspondence is alabastrum do not have different for a long time, each
Alabastrum launches only 3～4h (hour) just in cross from the sepal petal that ftractures, then through 1-2d (my god) after petal come off.Other is all
Such as bloom order, the flowering time etc. of every day of plant, sterile line does not has obvious difference with corresponding holding system.
(3) florescence of sterile line
Through early flowering season of wide handle mustard sterile line of continuous 7 generation backcross transformations compared with corresponding holding system, general postpone 2～
3d (my god), full-bloom stage and whole florescence postpone 3～5d accordingly (my god), the whole florescence continue 20-27d (my god), long reach 29d (my god) with
On, i.e. the florescence of sterile line than holding be long 5-6d (my god).
The fertility of wide handle mustard cytoplasm male sterility line
(1) fertility of stamen
1. the fertility under natural, ecological weather conditions: 2011～2015, each to wide handle mustard male sterility strain
While carrying out backcross transformation from generation to generation, centering menu strain has carried out bagging selfing and has measured, except the backcross generations strain of respective material
It is found to have beyond individual plant selfing fructifying index >=1 (made pottery and eliminated), finds no self-fertility phenomenon.2014～2015, the most right
The sterile lines such as " DJA, K6A, HK-1A, the K46A " after six generations that backcrossed and corresponding holding are that the solid situation of individual plant selfing is carried out
System is examined or check, 4 sterile strains either spring or the self-fertility index in summer is " 0 ", shows as the sterile of height
And fertility is highly stable；And keep tying up to different setting percentages in season and differ greatly, usually spring is solid solid higher than summer,
This may higher with summer temperature, plant nutrition body is little relevant.
2. the vitality of sterile line pollen: in March, 2014, to " DJA, K6A, HK-1A, HK-2A K46A, BKA " etc. 6
The pollen grain keeping system of sterile line and correspondence has carried out " TTC " dyeing microscopic examination to be observed.Result is: the pollen grain of sterile line is the most not
Coloring, and most pollen grain is that irregular roundness is spherical, transparent, partial pollen grain also has cracking, deficient phenomena, pollen grain edge in
Papillary；And keeping the pollen grain tinctorial yield being is 85-96%, shows as viable pollen and account for the overwhelming majority, pollen grain
Shape mostly be spheroidal or ellipse, opaque, full.
(2) fertility of gynoecium
2011～2015, the backcross transformation result of each generation showed, wide handle mustard sterile line (or sterile individual plant) female
Stamen stigma, after artificial pollination, not only can solid be set seeds, and offspring's growth promoter is normal, there is not reproduction isolation.Meanwhile, see
Examine the morphological characteristic of sterile line gynoecium, all do not find the difference having matter with corresponding holding system (or the male parent material that backcrosses).Thus
Proving, the Pistil Fertility of wide handle mustard male sterility line is the most normal.
S6, choose through backcrossing the transformation of seven generations and plant forms feature and the stable each sterile line of trait expression and corresponding keep
The young leaflet tablet tissue of system has carried out sterile line SSR molecular marker to be identified.That is: the 600 of the announcement of Cruciferae Brassica genus net are selected
To SSR primer sequence (being synthesized by Sheng Gong biological engineering company limited agency), use CTAB method to extract complete genome DNA, and divide
Do not carry out PCR reaction, electrophoresis and silver staining etc..Particularly as follows:
The 1-4 month in 2016 is 6 to the wide handle mustard male sterility line cultivated and holding (DJA and DJB, K6A and K6B,
HK-1A and HK-1B, K46A and K46B, HK-2A and HK-2B, BKA and BKB), select the 600 of the announcement of Cruciferae Brassica genus net
To SSR primer sequence, in Chongqing southeast Academy of Agricultural Sciences, carry out SSR molecular marker Screening and Identification, result such as Fig. 2 institute
Showing, in figure, 1-2 is sterile source JL154；3-5 is DJA-1-DJA-3；6-8 is K46A-1-K46A-3；9-10 be holding be DJ and
K46；11-13 is K6A-1--K6A-3；14-15 is BKA-1-BKA-2；16-17 is HK-1A and HK-2A；18-21 is for keeping system
K6、BK、HK-1、HK-2；22 is Maker.
The band that primer " KBJ416 " amplifies in sterile line and holding system is polymorphism.Wherein, swimming lane 1 is primer
The band that KBJ416 is amplified at tumorous stem mustard sterile line JL154-5A (on wide handle mustard sterile source)；Swimming lane 2-8,11-17 are for drawing
The band that thing " KBJ416 " amplifies on different wide handle mustard sterile lines；Swimming lane 9-10,18-21 are that primer " KBJ416 " is at wide handle
Mustard keeps fastening the band amplified.Meanwhile, band analysis shows, primer " KBJ416 " is at sterile source with through continuous backcross transformation
Different wide handle mustard cytoplasm male sterility lines on the band of amplification completely the same, and keeping with corresponding wide handle mustard is amplify
Banding pattern is presented with significant difference.
The above is only the preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For Yuan, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications, these improvements and modifications also should
It is considered as protection scope of the present invention.
Claims (1)
1. the selection of one kind wide handle mustard cytoplasm male sterility line, it is characterised in that comprise the steps:
Choose tumorous stem mustard cytoplasm male sterility line JL154-5A as female parent, and choose wide handle mustard Elite inbred as transformation father
Originally carrying out conventional hybridization and obtain F1, offspring selects typical case's individual plant and male parent continuous backcross 7 generation, it is thus achieved that sterile character is stable, economical
Wide handle mustard cytoplasm male sterility line and holding that character is good are.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107410015A (en) * | 2017-09-18 | 2017-12-01 | 华中农业大学 | A kind of selection of leafy mustard cytoplasmic male sterile line |
| CN113229139A (en) * | 2021-06-03 | 2021-08-10 | 重庆市渝东南农业科学院 | High-yield seed production method for fresh-eating leaf mustard |
| CN116326476A (en) * | 2021-12-10 | 2023-06-27 | 宁波市农业科学研究院 | Breeding method of processed dried mustard type mustard |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107410015A (en) * | 2017-09-18 | 2017-12-01 | 华中农业大学 | A kind of selection of leafy mustard cytoplasmic male sterile line |
| CN113229139A (en) * | 2021-06-03 | 2021-08-10 | 重庆市渝东南农业科学院 | High-yield seed production method for fresh-eating leaf mustard |
| CN116326476A (en) * | 2021-12-10 | 2023-06-27 | 宁波市农业科学研究院 | Breeding method of processed dried mustard type mustard |
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