CN106212269A - The method of the Herba Alii fistulosi new varieties of the selection-breeding high yield and high quality of resistance to storage - Google Patents
The method of the Herba Alii fistulosi new varieties of the selection-breeding high yield and high quality of resistance to storage Download PDFInfo
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Abstract
The present invention provides the method for the Herba Alii fistulosi new varieties of a kind of selection-breeding high yield and high quality of resistance to storage, with ' Folium Isatidis 1 ' Herba Alii fistulosi as material, sets up Herba Alii fistulosi sterile line breeding nursery, selection-breeding Herba Alii fistulosi male sterility line and holding system, sets up high-quality Herba Alii fistulosi male sterility line evaluation and screening system；Carry out individual plant selfing from the Herba Alii fistulosi resource of various trait, orthoselection is cultivated the male parent system stably isozygotied, and is carried out Identification of Comprehensive Characters, selects excellent male parent system；Utilizing the excellent male sterility line selected to hybridize with male parent system, preparing hybrid combines, by its offspring's keeping quality, yielding ability, the qualification of resistance, selecting Herba Alii fistulosi new varieties.The present invention establishes the evaluation of efficient male sterility breeding technique, screening system first, Innovation Germplasm resource, the resistance to storage of selection-breeding, high yield, disease-resistant, strong stress resistance, the triple crossing Herba Alii fistulosi new varieties of applicable cultivation of many seasons, the selection-breeding for high-quality Herba Alii fistulosi new varieties provides technical support.
Description
Technical field
The present invention relates to genetic breeding field, specifically, relate to the Herba Alii fistulosi new varieties of a kind of selection-breeding high yield and high quality of resistance to storage
Method.
Background technology
Herba Alii fistulosi is Liliaceae allium, originates from the Siberia of western part of China and the Soviet Union, wild Herba Alii fistulosi tame and docile in China
Changing and select, China is the major country cultivating Herba Alii fistulosi, is also the area of origin of Herba Alii fistulosi, by incoming Japan of Korea, and 1583
Year incoming Europe, 19th century incoming U.S., but Europe, U.S.'s cultivation are few, and world's Herba Alii fistulosi cultivated area 9,000,000 mu, China accounts for
8100000 mu.
Along with the development of Industrialization, Herba Alii fistulosi produces some problems existed and gradually displays: first, standing losses
Rate is too high, and overwhelming majority Folium Allii fistulosi loses, and particularly stores away in winter Herba Alii fistulosi and loses even more serious, accounts for through test measuring and calculating different cultivars Folium Allii fistulosi
55-65%；Secondly, improved seeds are few, and the kind in production mostly is conventional kind or its Improved lines and farmers' reserves seed for planting certainly, regularity
Difference, commodity is poor, yield poorly, resistance poor, storage property is the highest, and shelf life is short；Again, standardization level is low, lacks efficient
Corresponding cultivation techniques, it is impossible to given play to the high yield potential of kind, major part vegetable grower takes tradition planting patterns, and ability to accept is poor,
Relatively low to the Grasping level of scientic planting technology, it is difficult to realize the road of highly efficient and productive modern agricultural development.
China's Herba Alii fistulosi breeding uses a weak ring of traditional conventional herd breeding, the most always vegetable breeding mostly
Joint, lags behind other vegetable.Cause variet complexity owing to extensively introducing a fine variety, from numerous, breeding technique is lack of standardization etc., plant sexual involution, group
Body regularity difference and resistance are poor, can not meet production and market demand in terms of resistance and quality.Strengthen Herba Alii fistulosi breeding research
Become the task of top priority.Owing to Herba Alii fistulosi floral organ is little, single flower Setting percentage is low, and artificial emasculation cost is high, thus utilizes male sterility line
Preparing hybrid kind is one of maximally efficient approach.
Summary of the invention
The present invention is directed to problem present in Herba Alii fistulosi breeding and production, establish and set up efficient male sterility breeding skill
Art is evaluated, screening system, Innovation Germplasm resource, the resistance to storage of selection-breeding (Bulbus Allii Fistulosi Folium Allii fistulosi ratio is high), high yield, disease-resistant, strong stress resistance, be suitable for many
The triple crossing Herba Alii fistulosi new varieties that season cultivates are breeding objective.
In order to realize the object of the invention, the present invention provides the method for the Herba Alii fistulosi new varieties of a kind of selection-breeding high yield and high quality of resistance to storage,
Technology path is as follows:
What the compiling of the resource → qualification of resource, Innovation Germplasm resource → tri-were add for purification → set up sterile line and
Keep the evaluation of system, triage techniques system → assemble the Production of Large Fields qualification of Combination nova → F1, experiment in cultivation → obtain resistance to storage height
Produce the Herba Alii fistulosi new varieties of high-quality.
The method of the Herba Alii fistulosi new varieties of the selection-breeding high yield and high quality of resistance to storage of the present invention comprises the following steps:
S1, with ' Folium Isatidis 1 ' Herba Alii fistulosi as material, set up Herba Alii fistulosi sterile line breeding nursery, selection-breeding Herba Alii fistulosi male sterility line and guarantor
Hold and be, set up high-quality Herba Alii fistulosi male sterility line evaluation and screening system；
S2, from the Herba Alii fistulosi resource of various trait, carry out individual plant selfing, orthoselection cultivates the male parent system stably isozygotied, and
Carry out Identification of Comprehensive Characters, select excellent male parent system；
The male parent system hybridization that S3, the male sterility line utilizing S1 to obtain select with S2, preparing hybrid combines, by it
Offspring's keeping quality, yielding ability, the qualification of resistance, select Herba Alii fistulosi new varieties.
S1 is specific as follows:
S11, the selection-breeding of sterile plant
In No. 1 Herba Alii fistulosi multiplication fields of Folium Isatidis, screen Herba Alii fistulosi male sterility individual plant, be allowed to free pollination, according to Herba Alii fistulosi filial generation
Sterile plant fertility performance is different, filters out the excellent sterile strains that sterile rate is the highest；
S12, the selection-breeding of holding strain
In No. 1 Herba Alii fistulosi multiplication fields of Folium Isatidis, select fertility is normal, pollen is sufficient fine individual plant, sterile with selected by S11
The paired test cross of sterile plant that rate is the highest selfing, as keeping strain；
S13, sterile line, the pairing selection-breeding of holding system
From sterile plant, the seed of results is T system, and from fertile plant, the seed of results is S1 system, and sterile plant rate of selecting and remain is high
T system and the high S1 system of fertile plant rate, backcross respectively, selfing, plant division is collected backcross progeny BC1 and samsara male parent seed, is selected and remain
The individual plant that sterile plant rate is high and fertile plant rate is high, then carry out respectively backcrossing, selfing, by that analogy, until selecting high-quality Herba Alii fistulosi
Male sterility line and holding system.
Wherein, while S13 identifies fertility, male sterility line is utilized to use the mode preparing hybrid group of " the many mothers of father "
Close.
In S1, the criterion of male sterility line is as follows: 1. anther tapetum cell is done sth. in advance vacuolization and disintegrates, during disintegration
Between start from pollen mother cell period, be almost wholly absent period to monokaryon pollen grain, it is impossible to provide required for microspore development
Nutrient substance；2. the protoplasm concentration in tapetal cell is low, and without obvious many nuclear phenomena, the exception of this tapetal cell is also made
Become nutrient substance under-supply, cause pollen sterility；3. in monokaryon pollen grain to mature flower powder growth course, some flower
The connective cell fibrosis of medicine, hinders moisture and nutrient substance by vascular bundle to the conveying of pollen bag, thus causes pollen grain
Abortion.
S1 utilize primer combination nad5 and cox2 identify high-quality Herba Alii fistulosi male sterility line and keep system；
For the specific primer of PCR amplification nad5 gene to for (SEQ ID NO:1-2):
Nad5F:5 '-ATCTCTGAGGATCCGCATAGC-3 '
Nad5R:5 '-ACATCGTAACCTGTCTCGC-3 '
For the specific primer of PCR amplification cox2 gene to for (SEQ ID NO:3-4):
Cox2F:5 '-CGATTCCAGAAGATGATCCAG-3 '
Cox2R:5 '-GGATACCCGAGAACCATAATC-3 '
Aforesaid method, the high-quality Herba Alii fistulosi male sterility line selected in S1 is ' 08 blue or green 601A '.
Aforesaid method, the high-quality Herba Alii fistulosi male sterility line selected in S1 keeps system for ' 08 blue or green 601B '.
Primer can amplify the band of about 1180bp size to nad5 in 08 blue or green 601B keeps being, and 08 blue or green 601A is not
Educate be in do not have.Primer can keep amplifying in system about 1750bp and about 380bp two band at 08 blue or green 601B to cox2, and 08
Blue or green 601A sterile line does not has.
Aforesaid method, Herba Alii fistulosi resource described in S2 includes ' Zongzheng Herba Alii fistulosi king ' etc..
Aforesaid method, also includes in S3 that using packet wheel to join method carries out the mensuration of combining ability: be classified as by male sterility line
Maternal group, same male parent and each male sterility line group gas-mixing hybridization, same male sterility line is miscellaneous as maternal and multiple male parent combos
Hand over.
The present invention establishes the evaluation of efficient male sterility breeding technique, screening system, Innovation Germplasm resource, selection-breeding first
Resistance to storage (Bulbus Allii Fistulosi Folium Allii fistulosi ratio is high), high yield, disease-resistant, strong stress resistance, the triple crossing Herba Alii fistulosi new varieties of applicable cultivation of many seasons, for high-quality
The selection-breeding of Herba Alii fistulosi new varieties provides technical support.
Accompanying drawing explanation
Fig. 1 is the stamen form and pollen vitality that in present pre-ferred embodiments, sterile line and holding are；Wherein, A:a
For sterile line stamen, b be holding be stamen；B:a is the flower pesticide that sterile line is shrivelled, b be holding be flower pesticide.
Fig. 2 is to keep system and the mature pollen grain form of sterile line and vigor in present pre-ferred embodiments；Wherein, A-B:
Holding is pollen morphology and vigor；C: sterile line pollen morphology and vigor.
Fig. 3 is that in present pre-ferred embodiments, sterile line and holding are the microstructure of flower pesticide；Wherein, A: sterile line pollen
Blast cell is in early days (400 ×)；In B: the sterile line pollen mother cell later stage, arrow shows tapetal cell vacuolization (400 ×)；C: keep
Be pollen mother cell to the subtrahend first division phase, arrow shows normotrophic tapetal cell (400 ×)；D: sterile line subtrahend
Division stage, arrow shows the tapetal cell (400 ×) disintegrated；E: holding is reduction division phase, and arrow shows normal carpet
Confluent monolayer cells (400 ×)；F-H: sterile line children's tender pollen grain phase, F, G (400 ×), H (200 ×)；I-K: holding is young tender pollen grain
Phase (400 ×)；L: sterile line mature flower powder phase (400 ×)；M: sterile line connective cell fibrosis (100 ×), lower left is
Fibrosis connective cell (400 ×) amplified；The empty flat pollen grain (400 ×) of N: sterile line；O: keep being tied to form ripe pollen grain (400
×)。
Fig. 4 is different primers amplification on the 08 blue or green 601B of blue or green 601A and 08 in present pre-ferred embodiments；Wherein,
①orfA501②atp1③atp6④nad5⑤cox2⑥cox3⑦rrn26。
Fig. 5 is the technology road figure line of the Herba Alii fistulosi new varieties of the selection-breeding high yield and high quality of resistance to storage in present pre-ferred embodiments.
Fig. 6 is that in present pre-ferred embodiments, sterile line keeps is Breeding Process figure.
Detailed description of the invention
Following example are used for illustrating the present invention, but are not limited to the scope of the present invention.If not specializing, embodiment
All according to conventional laboratory conditions, as Sambrook equimolecular Cloning: A Laboratory Manual (Sambrook J&Russell DW,
Molecular Cloning:a Laboratory Manual, 2001), or the condition according to manufacturer's description suggestion.
The method of the Herba Alii fistulosi new varieties of the embodiment selection-breeding high yield and high quality of resistance to storage
Material based on No. 1 Herba Alii fistulosi of Folium Isatidis, sets up Herba Alii fistulosi sterile line breeding nursery, innovates Herba Alii fistulosi germ plasm resource, and selection-breeding is big
Herba Alii fistulosi male sterility line and holding system, set up high-quality Herba Alii fistulosi male sterility line evaluation and screening system.From the difference collected both at home and abroad
Carry out individual plant selfing in character Herba Alii fistulosi resource, orthoselection is cultivated the male parent system stably isozygotied, and is carried out Identification of Comprehensive Characters, choosing
Bring out excellent male parent system.According to breeding objective, utilizing sterile line to hybridize with male parent system, preparing hybrid combines.By to its offspring
Keeping quality compares, yielding ability compares, resistance etc. is identified, identifies excellent combination, and then selects and meet the big of breeding objective
Herba Alii fistulosi new varieties (Fig. 5).
(1) breeding technique research
Owing to Herba Alii fistulosi floral organ is little, single flower Setting percentage is low, and artificial emasculation cost is high, thus utilizes male sterility line preparation miscellaneous
Friendship kind is maximally efficient approach.And the evaluation of male sterility line and rapid screening can greatly reduce workload, improve breeding
Efficiency.
The present invention, in implementation process, for the Herba Alii fistulosi germ plasm resource of project team, constructs and acquires cytology again from form
To molecular marker assisted selection again to male sterility line combining ability test evaluation and screening system.
1, morphology
(1) plant forms
Sterile line 08 blue or green 601A: plant height 105cm, Bulbus Allii Fistulosi 48.2cm, thick 3.1cm, upright blade, dark green leaf color, blade face wax
Powder is many, and field growing is healthy and strong.High anti-Onion Purple Blotch, downy mildew, virosis.Degenerate slow, resistance.There is excellent high yield excellent
Matter character.
(2) flower pesticide form
The blue or green 601A stamen of Herba Alii fistulosi male sterility line 08 has normal flower pesticide and filigree, flower pesticide profile to show as shrivelled not opening
Splitting, WUHUAFEN grain sheds (Fig. 1).This feature is suitable for the preliminary field Rapid identification of this male sterility line.
2, cytology
(1) mensuration of pollen grain activity
This sterile line pollen bag does not ftractures, and flower pesticide is contained within pollen grain, but pollen grain inactive (Fig. 2), belong to pollen and lose
Educating the sterile line of type, this feature is conducive to field Rapid identification male sterility.
(2) cytologic characteristic of male sterility line anther development
In addition to identifying in morphology, by sterile line and system is kept to carry out paired observation from cytological angle.No
Educating the anther structure feature being as follows: 1. anther tapetum cell is done sth. in advance vacuolization and disintegrates, it is female thin that removal time starts from pollen
Born of the same parents period, almost it is wholly absent period to monokaryon pollen grain, it is impossible to provide required nutrient substance for microspore development；2. carpet
Protoplasm concentration in confluent monolayer cells is low, and without obvious many nuclear phenomena, the exception of this tapetal cell also results in nutrient substance supply
Deficiency, causes pollen sterility；3., in monokaryon pollen grain to mature flower powder growth course, the connective cell of some flower pesticide is fine
Dimensionization, hinders moisture and nutrient substance by vascular bundle to the conveying of pollen bag, thus causes pollen sterility.Sterile line and guarantor
Holding is anther development comparative structure figure (Fig. 3).
3, the blue or green 601A sterile line of molecular marker assisted selection 08 and 08 blue or green 601B keep system
Forefathers are it is believed that the generation of cytoplasmic male sterile line (CMS), with the intermolecular of mitochondrial genome or molecule
In frequently restructuring and the mosaic gene that formed is relevant.
The present invention according to the portion gene of mitochondrial genome (nad5, cox2, cox3, atp1, atp6, rrn26,
OrfA501, SCS13, SCS200) and the method for reference Wang Chan, Wu Haitao etc., synthetic primer pair, expand through PCR, screen 2
To primer, (nad5 and cox2, the specific primer of amplification nad5 gene, to as shown in SEQ ID NO:1-2, expands cox2 gene
Specific primer to as shown in SEQ ID NO:3-4) can identify that 08 blue or green 601A sterile line and 08 blue or green 601B keep system.
Nad5 (nadh dehydrogenase 5) can amplify the band of about 1180bp in 08 blue or green 601B keeps being, and does not has in sterile line.
Cox2 (cytochrome c oxidase 2) can keep amplifying about 1750bp and about 380bp two band in system at 08 blue or green 601B, and
08 blue or green 601A sterile line do not have (Fig. 4).
Utilize the two primer can keep system with the blue or green 601A sterile line of Rapid identification 08 and 08 blue or green 601B.
(2) collection of germ plasm resource and qualification
Collect different types of domestic and international Herba Alii fistulosi resource 26 parts, and plant together with original resource, identify its yielding ability,
Disease resistance, resistance etc., filter out excellent breeding material.Carry out classifying, arrange, preserve, and for breeding objective, by these
Resource is purified seed selection and mutative material retains innovation.Utilize the plot planting Herba Alii fistulosi for many years to create Herba Alii fistulosi Infected Field, adopt
With controlling the method that worm is not cured the disease, material and disease-resistant individual plant to excellent resistant effectively select.
For breeding objective, the material collected is screened and identified.Different variety sources is planted in consistent
Under cultivation condition, it is allowed to the difference fully showed each other.In order to make the Horticultural Characters of different cultivars group fully manifest, we are carried out
Subregion is planted, and takes corresponding cultivation technique measure.About 75,3 ridgings of long white type kind line-spacing, short white and drumsticks class
About 60,2 ridgings of type kind line-spacing.From differences such as the plant of Herba Alii fistulosi, leaf, cauloid, flowers during whole plant strain growth
Growth stage repeatedly carry out screening, identifying.
The major product of Herba Alii fistulosi is Bulbus Allii Fistulosi, and it is covered among ridging in field, it is impossible to complete detection, strain commodity are gathered
After, carry out field species test qualification.
(3) breeding of new variety
1, the selection-breeding of Herba Alii fistulosi male sterility line
The selection-breeding of 1.1 sterile plants
Within 2006, in No. 1 Herba Alii fistulosi multiplication fields of Folium Isatidis, find 12 strain Herba Alii fistulosi male sterility individual plants, list numbered 06 respectively
Sesbania cannabina (Retz.) Pers. CA-1,06 Sesbania cannabina (Retz.) Pers. CA-2 ... 06 Sesbania cannabina (Retz.) Pers. CA-12, be allowed to free pollination, after spontaneous pollination, gathers in the crops guarantor the most respectively
Deposit.The same year is by individual plant nursery mid-July of results, open country net canopy field planting in October.The investigation in florescence of the first tenday period of a month in May, 2007 Herba Alii fistulosi
Finding, selected 12 strain Herba Alii fistulosi sterile plant fertility performance are different, and wherein 06 Sesbania cannabina (Retz.) Pers. CA-2 shows sterile rate and is up to 95%, other quilt
Eliminate.
1.2 selection-breeding keeping strain
From Herba Alii fistulosi seed farm, within 2006, select 12 fine individual plants that strain fertility is normal, pollen is sufficient, extract open flower,
Bagging selfing, gathers in the crops numbering respectively.In July, 2006 is sowed, field planting in October open country net canopy.In May, 2007, Herba Alii fistulosi selected when blooming excellent
Good individual plant, extracts open flower and lists bagging, and the selected paired test cross of sterile plant selfing, as keeping strain.
1.3 sterile lines, the pairing selection-breeding of holding system
Within 2007, selecting 50 strain excellent sterile strains to extract open flower from 06 Sesbania cannabina (Retz.) Pers. CA-2, bagging of listing, from institute in 2007
12 strains of choosing can be educated in individual plant offspring, selects the educated individual plant that pollen amount is sufficient, extracts open flower, bagging of listing, 3-4 days
After, swipe pollen pollinates (test cross T) to selected sterile plant in pairs, every day 1 time, individual plant results seed after seed maturity, never
Educating the seed of results in strain is T system, and from fertile plant, the seed of results is S1 system.
The seed of results is divided two parts to process by the same year: a part of nursery in July, a part of nursery in October.Nursery in July
Herba Alii fistulosi October T system and S1 be tied to form field planting made Fertility identification to open country net canopy, May next year when blooming, investigate T respectively
The S1 system that system and S1 system is sterile and fertile plant rate, T system that sterile plant rate of selecting and remain is high and fertile plant rate are high, is selected in strain and respectively selects 50
Strain, backcrosses, selfing respectively, and backcross combination (BC1) and samsara male parent seed (S2) are collected in plant division.The Herba Alii fistulosi of nursery in October is in 2008
June in year is by high in July, 2007 nursery fertility in May next year investigation sterile plant rate and that fertile plant rate is high pair-wise combination field planting land for growing field crops
In, remaining is eliminated.Carry out field species test in November, 2008, strain is identified, eliminates the strain of field growing inferior position.And from pairs
Big maternal plant in select the excellent or sterile plant of variation and keep strain group to and field planting open country net canopy.In May, 2009 makees fertility
Identify and select.Time (Fig. 6) by that analogy later.
Within 2012, select Anther degeneration WUHUAFEN at breeding nursery, sterile rate close to 100%, upright blade, leaf color green or
The excellent Herba Alii fistulosi male sterility line 4 of the Comprehensive Traits such as dark green, disease-resistant, resistance is right, ' 08 blue or green 601A ', ' 08 blue or green 353A ', ' 04 blue or green
Standby C3A ', ' 04 blue or green standby C9A ' and the holding matched are 08 blue or green 601B ', ' 08 blue or green 353B ', ' 04 blue or green standby C3B ', ' 04 blue or green standby
C9B ' (table 1, table 2).
Table 1 sterile line, holding are major traits application form
The blue or green sterile rate of 601A of table 2 Herba Alii fistulosi sterile line 08 and corresponding the educated rate application form keeping system
Utilize male sterility line to use the method preparing hybrid combination of " the many mothers of father " while identifying fertility, and carry out miscellaneous
Hand over the qualification of combination and the selectable little area production of hybrid seeds, arrange many ground multiple spot adaptability to identify.
The feature of 1.4 Herba Alii fistulosi male sterility lines
Sterile line 08 blue or green 601A: plant height 105cm, Bulbus Allii Fistulosi 48.2cm, thick 3.1cm, upright blade, dark green leaf color, blade face wax
Powder is many, and field growing is healthy and strong.High anti-Onion Purple Blotch, downy mildew, virosis.Degenerate slow, resistance.
2, the selection-breeding of Herba Alii fistulosi male parent system
2.1 selection-breeding are passed through
The selection-breeding starting Herba Alii fistulosi male parent system in 2006.Infected Field Disease Resistance Identification and individual plant orthoselection is used to combine
Method.According to breeding objective from the domestic and international Herba Alii fistulosi resource of breeding nursery and introduction, select 20 excellent strain (selfings of character
More than 1 generation), each selection 10 strains, field planting in October land for growing field crops open country net canopy, when May blooms, artificial bagging pollination self, individual plant
Results, the seed nursery in October after results, field planting in June, 07 land for growing field crops.More than each strain field planting 200 strain, in November, 07
Part land for growing field crops species test, qualification, strictly select, and superseded character does not meets the strain 78 of breeding objective, remains 22 and is selected in, and by this
22 strains strictly rogue select excellent, from the shape of plant, Bulbus Allii Fistulosi length, thickness, disease resistance, regularity, the elasticity etc. of Bulbus Allii Fistulosi
Selecting excellent in excellent, each strain stays 8-10 strain, in field planting in March, 10 open country net canopy.When May, Herba Alii fistulosi bloomed, artificial bagging
Pollination self, gathers in the crops after seed maturity respectively.Seed now have passed through the selection-breeding in 3 generations more than, and the trend having is stable, this
Time in order to accelerate Breeding progress, improve its vitality (often substantially having decay for selfing vitality) and use little maternal plant to add generation
Breeding.The seed of results is divided two parts, part nursery in July, 11, a part of nursery in October, the kind of nursery in July
Son, field planting in November, 11 open country net canopy, each strain when in May, 12 blooms selects 6 strains to carry out bagging pollination, nursery in October
Field planting in June, 12 land for growing field crops, November field test, according to the growing way of plant, the number of blade, cauloid length, thickness, the degree of packing,
The character directive breedings such as resistance, excellent in select excellent, cultivate and meet the excellent height of breeding objective Comprehensive Traits for 9 (tables of selfing line
3)。
Table 3 Inbred Lines on Main application form
The feature of 2.2 male parent systems
Male parent 08-21-3-7: source kind Zongzheng, Shandong Herba Alii fistulosi king, plant height 115cm, the long 59.6cm of Bulbus Allii Fistulosi, white thick 3.3cm,
Bulbus Allii Fistulosi consolidation；5-6 piece of tubulose leaf, leaf green, the long 58cm of leaf.Mouthfeel is crisp sweet, fast growth, weatherable, and resistance to overhead flooding injury is good.
3, the selection-breeding of Herba Alii fistulosi new varieties is hybridized
3.1 selection-breeding are passed through
Within 2012, while carrying out sterile line, keeping system and male parent system to add generation purification screening, use packet wheel to join method and enter
Having gone the mensuration of combining ability, will be classified as maternal group by male sterility line, selfing line is classified as male parent group, same male parent with each male not
Educating is combo, and same male sterility line is made maternal and multiple male parent selfing line and assembled, and filters out the hybridization group meeting breeding objective
Close.
36 cross combinations to being prepared in 2013 carry out Preliminary Identification, the group of numbered 13-26 in this tested field
Conjunction is had outstanding performance.This combination female parent is the blue or green 601A of Herba Alii fistulosi male sterility line 08, and male parent system is high for selfing line 08-21-3-7, fixed
Entitled ' blue or green miscellaneous No. 3 '.This uniform quality degree is high, blade is dark green, blade face waxiness thickness, upright blade, high yield, disease-resistant, resistance
Prominent, strong adaptability.
3.2 selection-breeding results
3.2.1 keeping quality
The qualification of keeping quality: same identifies field kind, by unified standard, respectively to, blue or green miscellaneous No. 3 and cold rich huge Herba Alii fistulosi respectively choose
Uniform 15 strains, weigh after arrangement, after surveying Bulbus Allii Fistulosi Folium Allii fistulosi ratio, are placed under in the shade natural conditions by sample, bury Bulbus Allii Fistulosi with soil, point
Weigh respectively for twice, computed losses rate (table 4).
Blue or green miscellaneous No. 3 Bulbus Allii Fistulosi Folium Allii fistulosi comparative control are high by 10%.
Table 4 Bulbus Allii Fistulosi Folium Allii fistulosi is than test result (repeating average three times)
In storing away in winter test, the loss of blue or green miscellaneous No. 3 is 27.1, and check variety is 44.7%.Significantly lower than check variety.(table
5)
Table 5 Herba Alii fistulosi winter storage test experimental result (repeats average three times)
Yielding ability is good: on the one hand 2014-2015 carries out variety comparative test in this locality, on the other hand in Zanhuang, Shijiazhuang, Handan
Ci County, Dancheng, Xinxiang, Henan carry out multiple spot pilot production.Nursery on October 3, field planting on June 12 in next year, same plot, identical pipe
Reason, check variety is Zhangqiu in Shandong province's green onion breed-cold Feng Jucong.' blue or green miscellaneous No. 3 ' Herba Alii fistulosi is at plant height, horizontal stem, vertical stem, Weight per plant
Amount is significantly better than that check variety.Variety comparative test sets 3 repetitions, random alignment, result show blue or green miscellaneous No. 3 more continuous than check variety
2 increase production every year, and amount of increase in production is 19.7-21.1% (table 6), and pilot production uses random alignment, takes the mean at (every 15 at 3
), ' blue or green miscellaneous No. 3 ' all ratio comparison volume increase 3 testing sites in continuous 2 years, amount of increase in production is 17.8-21.2%.(table 7).Produce
Starting popularization while test, crop cultivation speciality cooperative society is consistent with plantation family thinks that this Herba Alii fistulosi regularity is high, grows vigorous,
Folium Allii fistulosi is dark green, disease-resistant, strong stress resistance, than local main breed increase yield significantly.
The blue or green miscellaneous No. 3 Herba Alii fistulosi variety comparative test results of table 6
The blue or green miscellaneous No. 3 Herba Alii fistulosi pilot production yield of table 7
3.2.2 disease resistance is strong
In 2014-2015 Herba Alii fistulosi variety comparative test, the disease resistance to ' blue or green miscellaneous No. 3 ' is investigated, and its result is: blue or green miscellaneous 3
Number downy mildew, anaphylactoid purpura and the sickness rate of virosis and disease index are trembled with fear Feng Jucong significantly lower than check variety, show superpower
Premunition, high anti-big onion mildew, anaphylactoid purpura and virosis (table 8).
Table 8 Herba Alii fistulosi disease resistance statistical result
Note: investigation strain number is 100 strains.
The most anti-fall, resistance to overhead flooding injury good
Investigating under windy, rainy weather condition in 2014, investigation place is village of thousand, Tu Men township, Zanhuang County.In Herba Alii fistulosi
Plot is concentrated in plantation, chooses medium management level plot, randomization, and every piece of ground takes 3 points, and every takes 100 strains.Result shows
(table 9): the lodging resistance relatively check variety of ' blue or green miscellaneous No. 3 '--cold rich huge Herba Alii fistulosi lodging rate is low by 10%.
Under conditions of rainy can normal growth, the multiple diseases generation very small amount such as downy mildew, anaphylactoid purpura, virosis.Comparison
Kind tremble with fear rich huge onion mildew, anaphylactoid purpura, virosis etc. sick occur more serious.
The blue or green miscellaneous No. 3 lodging resistance application forms of table 9
3.2.4 strong adaptability
' blue or green miscellaneous No. 3 ' as spring Herba Alii fistulosi, summer Herba Alii fistulosi, volt Herba Alii fistulosi, autumn scallion and winter storage Herba Alii fistulosi cultivation, and can possess as winter storage Herba Alii fistulosi
Well-grown, resistance highlights, resistance to storage, yield high.
3.2.5 the feature of new varieties
This uniform quality degree is high, and plant is tall and big, and growth potential is strong, fast growth；Upright blade, dark green leaf color, tubulose leaf 6
Piece, blade face wax powder is thick, and wind resistance, lodging resistant force are strong.Blade upper punch or the most tiltedly stretch, mesophyll is thicker；Plant height 150cm, the long 55-of Bulbus Allii Fistulosi
70cm, thick about 4cm, long about the 70cm of leaf, Weight per plant 0.45-0.55kg, Bulbus Allii Fistulosi Folium Allii fistulosi compares 14:11；Quality is tight, tender and crisp,
Taste is sweet, average yield per mu 7502.6kg, than comparison volume increase 19.6%；Cold-resistant waterlogging, high anti-anaphylactoid purpura, virosis, downy mildew, adapt to
Property strong, cultivation area is wide, is suitable for four seasons cultivation, stores away in winter the preferable kind of Herba Alii fistulosi cultivation especially.
Although, the present invention is described in detail the most with a general description of the specific embodiments, but
On the basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements without departing from theon the basis of the spirit of the present invention, belong to the scope of protection of present invention.
List of references
Wang Chan. the microexamination of Herba Alii fistulosi male sterility line and relevant DNA fragmentation screening [D]. University Of Shanxi's master's degree opinion
Literary composition, 2013 Wu's great waves, Wang Jianjun, Hou Xilin, Liu Hongjiong, Ma Rongli, Ma Jingfan. two Bulbus Allii Cepae male sterility line cytoplasm types
Qualification with analyze [J]. gardening journal, 2009,36 (5): 723-726.
Claims (8)
1. the method for the Herba Alii fistulosi new varieties of the selection-breeding high yield and high quality of resistance to storage, it is characterised in that comprise the following steps:
S1, with ' Folium Isatidis 1 ' Herba Alii fistulosi as material, set up Herba Alii fistulosi sterile line breeding nursery, selection-breeding Herba Alii fistulosi male sterility line and keep system,
Set up high-quality Herba Alii fistulosi male sterility line evaluation and screening system；
S2, from the Herba Alii fistulosi resource of various trait, carry out individual plant selfing, orthoselection is cultivated the male parent system stably isozygotied, and is carried out
Identification of Comprehensive Characters, selects excellent male parent system；
The male parent system hybridization that S3, the male sterility line utilizing S1 to obtain select with S2, preparing hybrid combines, by its offspring
Keeping quality, yielding ability, the qualification of resistance, select Herba Alii fistulosi new varieties.
Method the most according to claim 1, it is characterised in that S1 is specific as follows:
S11, the selection-breeding of sterile plant
In No. 1 Herba Alii fistulosi multiplication fields of Folium Isatidis, screen Herba Alii fistulosi male sterility individual plant, be allowed to free pollination, sterile according to Herba Alii fistulosi filial generation
Strain fertility performance is different, filters out the excellent sterile strains that sterile rate is the highest；
S12, the selection-breeding of holding strain
In No. 1 Herba Alii fistulosi multiplication fields of Folium Isatidis, select fertility is normal, pollen is sufficient fine individual plant, with the sterile rate selected by S11
The paired test cross of high sterile plant selfing, as keeping strain；
S13, sterile line, the pairing selection-breeding of holding system
From sterile plant, the seed of results is T system, and from fertile plant, the seed of results is S1 system, the T system that sterile plant rate of selecting and remain is high
The S1 system high with fertile plant rate, backcrosses, selfing respectively, and backcross progeny BC1 and samsara male parent seed, sterile plant of selecting and remain are collected in plant division
The individual plant that rate is high and fertile plant rate is high, then carry out respectively backcrossing, selfing, by that analogy, until it is male not to select high-quality Herba Alii fistulosi
Educate and be and keep system.
Method the most according to claim 2, it is characterised in that while identifying fertility in S13, utilize male sterility line to adopt
Combine with the mode preparing hybrid of " the many mothers of father ".
4. according to the method described in any one of claim 1-3, it is characterised in that in S1, the criterion of male sterility line is such as
Under: 1. anther tapetum cell is done sth. in advance vacuolization and disintegrates, and removal time starts from pollen mother cell period, during to monokaryon pollen grain
Phase is almost wholly absent；2. the protoplasm concentration in tapetal cell is low, without obvious many nuclear phenomena；3. arrive at monokaryon pollen grain
In mature flower powder growth course, the connective cell fibrosis of flower pesticide.
5. according to the method described in any one of claim 1-4, it is characterised in that S1 utilizes primer combination nad5 and cox2 mirror
Determine high-quality Herba Alii fistulosi male sterility line and keep system；
For PCR amplification nad5 gene specific primer to for:
Nad5F:5 '-ATCTCTGAGGATCCGCATAGC-3 '
Nad5R:5 '-ACATCGTAACCTGTCTCGC-3 '
For PCR amplification cox2 gene specific primer to for:
Cox2F:5 '-CGATTCCAGAAGATGATCCAG-3 '
Cox2R:5 '-GGATACCCGAGAACCATAATC-3 '.
6. according to the method described in any one of claim 1-5, it is characterised in that the high-quality Herba Alii fistulosi male sterility selected in S1
System is ' 08 blue or green 601A '.
7. according to the method described in any one of claim 1-6, it is characterised in that Herba Alii fistulosi resource described in S2 includes ' Zongzheng Herba Alii fistulosi
King '.
8. according to the method described in any one of claim 1-7, it is characterised in that S3 also including, using packet wheel to join method is carried out
The mensuration of combining ability: male sterility line is classified as maternal group, same male parent and each male sterility line group gas-mixing hybridization, same male not
Educate is as maternal and multiple male parent group gas-mixing hybridizations.
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| CN104946633A (en) * | 2015-06-02 | 2015-09-30 | 山东省农业科学院蔬菜花卉研究所 | dCAPS label for identifying cytoplasmic fertility of welsh onion and application thereof |
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| CN111670802B (en) * | 2020-07-06 | 2021-11-30 | 石家庄市农林科学研究院 | Breeding method of long-white chicken leg type green Chinese onions |
| CN111820094A (en) * | 2020-07-24 | 2020-10-27 | 石家庄市农林科学研究院 | Planting method for shortening breeding period of green Chinese onions |
| CN116114590A (en) * | 2022-11-28 | 2023-05-16 | 山东省种子有限公司 | Method for improving probability of green Chinese onion male sterile line and application thereof |
| CN116114590B (en) * | 2022-11-28 | 2024-04-16 | 山东省种子有限公司 | Method for improving probability of green Chinese onion male sterile line and application thereof |
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