CN105359963B - Method for cultivating easy-coloring apple variety - Google Patents
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Abstract
The invention discloses a method for cultivating an easy-coloring apple variety. An apple breeding method provided by the invention comprises the following steps: (1) hybridizing an apple cultivar A and an apple cultivar B to obtain hybrid seeds, wherein the apple cultivar A has genotype of R6R6 homozygotic type or R1R6 heterozygous type, and the apple cultivar B has genotype of R1R1 homozygotic type; (2) sowing and seedling the hybrid seeds obtained in the step (1) to obtain seedlings; and (3) transplanting the seedlings obtained in the step (2) to the field, skipping bagging after fruiting, and screening to obtain plants with specific characters of the anthocyanin content of fruit pulp higher than 0.010mg/g FW. The method provided by the invention can foster apple variety with excellent comprehensive quality and easy coloring property, and has great significance to the establishment of bagging-free apple cultivating system in our country, and to cost reduction and efficiency improvement. The invention has great value to apple breeding.
Description
Technical field
The present invention relates to a kind of easy coloring apple variety cultivating method.
Background technology
Rural Poverty Population realizes that it is the key for building up well-off society comprehensively to shake off poverty, most difficult during being also China " 13 "
One of huge goal task.And it is economical descending, agricultural product price is persistently dropped, and " yellow case " policy of agriculture subsidy is also arrived substantially
" ceiling ", " floor " constantly lifting of production cost；Under the Double Pressures on " ceiling " and " floor ", Apple Industry can
Continue high-efficient development and Practices Sustainable Increasing Income of Farmers, one will expand apple consumption market, two reduces costs and price, improve work production
Efficiency and labor productivity, cost-saving synergistic.
Apple Bags cultivation, in existing more than the 10 years history of China's large-area applications, is China's high quality apple production at this stage
A key technology, to improve fruit appearance quality, control disease pest harm, reduce residues of pesticides played important function.But
In recent years, with the increase year by year of labor cost, by fruit bagging, pluck the orchard recruitment cost that bag causes and turned into current apple
Fruit produces main input cost；Finding shows, about average 5000 yuan/mu of production cost in current China apple orchard,
Wherein bagging, to pluck the cost that bag causes be 2600 yuan, accounts for more than the 50% of total production cost；The current apple orchard gross area of China
37000000 mu, bagging is put into every year, the costly up to 96,200,000,000 yuan of bag is plucked；And it is the soluble solid content of Apple with Fruit-bagged Treatment, total
Sugared content reduction, acidity is improved, and Studies of The Aromatic Substances content is reduced, and local flavor is thin out；Peeling hardness reduction, fruit face cuticula, wax coat become
Thin, respiratory metabolism and moisture loss aggravate, and storage performance is deteriorated, and apple interior quality declines；Meanwhile, Apple with Fruit-bagged Treatment bitter pit,
Diplostomiasis, red spot disease etc. occur serious.Therefore, how to carry out apple to be cultivated without bagization, reduce the orchard amount of labour used and labour cost,
Fruit interior quality is improved, into the problem that orchard worker is in the urgent need to address.It is not simple not bagging without bag cultivation, but
Modern wide row high level cadre cultivation mode is set up, improve orchard ventilation and penetrating light condition, reduce pest and disease damage occurrence index, increase by studying
Organic fertilizer puts into and reduces a series of measures such as chemical fertilizer (N fertilizer) usage amount, makes the apple of not bagging in fruit face finish, coloring etc.
Aspect reaches or basically reaches the standard of Apple with Fruit-bagged Treatment.It is the pass for exempting from pocket culture wherein to cultivate the strong apple variety of colorability
One of key measure.
Apple storage property is good, and supply cycle is long, fruit contain higher proportion, human body be easier to absorb free polyphenol
(flavonoids), with the effect such as anti-oxidant, antitumor, prevention cardiovascular and cerebrovascular disease and protect liver well, healthy nutritive value is high,
There is the good reputation of " one day apple, doctor is away from me ", considerable country is all classified as major consumers fruit and big in the world
Push and recommend；Anthocyanin is the important component of flavonoids, is the constituent element of nutritional quality not only with healthy nutritive value, and
And be the mcintosh kind Red color trait development main matter that i.e. exterior quality is formed.Therefore, apple anthocyanin biosynthesis
Research receive extensive concern, achieve many impressive progresses.
The content of the invention
It is an object of the invention to provide a kind of easy coloring apple variety cultivating method.
The Apple breeding method that the present invention is provided, comprises the following steps：
(1) apple variety A and apple variety B are hybridized, is obtained hybrid seed；The apple variety A is genotype
For R6R6 is homozygous or the apple variety of R1R6 heterozygous；The apple variety B is that genotype is the homozygous apple product of R1R1
Kind；
(2) the hybrid seed sowing and nursery for obtaining step (1), obtain seedling；
(3) transplantation of seedlings to crop field of growing directly from seeds for obtaining step (2), bagging is not carried out after it grows fruit to fruit, sieve
Choosing obtains the plant with specific trait；The specific trait is：The pulp Anthocyanin Content of fruit is 0.010mgg-1FW with
On；
The method for judging the genotype of apple variety is as follows：
Genomic DNA with apple to be measured as template, using sequence table sequence 1 shown in single strand dna and sequence
The primer pair of the single strand dna composition shown in the sequence 2 of table enters performing PCR amplification, then by following standard interpretation genotype：Such as
Fruit pcr amplification product is a kind of and is 500bp, and the genotype of apple to be measured is that R6R6 is homozygous；If pcr amplification product is one
Kind and be 382bp, the genotype of apple to be measured is homozygous for R1R1；If pcr amplification product be two kinds and for 500bp and
382bp, the genotype of apple to be measured is R1R6 heterozygous.
The specific trait is concretely：The fruit face of fruit is bright and clean, light, and the pulp Anthocyanin Content of fruit is
0.010mg·g-1More than FW.
The genotype is that the homozygous apples of R6R6 can be purplish red No. 1, purplish red No. 2 or purplish red No. 3.
The genotype can be red crisp No. 1 for the apple of R1R6 heterozygous.
In the step (2), before carrying out the sowing, first the hybrid seed is carried out 1-3 DEG C of Stratificated treatment to break
Dormancy.The concretely 60 days time of the Stratificated treatment.
In the step (2), the condition for carrying out the nursery is：25 DEG C, daily illumination 12 hours, intensity of illumination
3000lx, it is every since seed is sprouted to pour within 7-10 days one time of nutrition liquid.
In the step (2), the condition for carrying out the nursery is specially：The hybrid seed is seeded in equipped with nursery base
Nutritive cube (each nutritive cube the sows 3-5 seed) culture of matter is highly that lignifying is (logical at 8-15cm and root neck to seedling
It is often sowing 2-3 month), then growing directly from seeds, to the new nutritive cube equipped with seedling medium, (each nutritive cube transplants 1 for transplantation of seedlings
Strain) cultivated；The condition of the culture is：25 DEG C, daily illumination 12 hours (intensity of illumination 3000lx), sprout from seed and open
Beginning often, pour within 7-10 days one time of nutrition liquid (is diluted with water to 10 times of volume as nutrition by a great number of elements mother liquor of MS minimal mediums
Liquid), every time each nutritive cube pour 40-50ml.
In the step (3), the method for carrying out cultivation management in the big Tanaka is as follows：Drafting the crop field of plant seedling
In every mu apply organic fertilizer 6000kg and water heavy real, apply organic fertilizer 6000kg again simultaneously in big every mu of Tanaka after field planting seedling
Water in time.The cattle manure that the organic fertilizer concretely fully becomes thoroughly decomposed.
In methods described, transplant the 4th year, the trunk to seedling at 20cm from the ground carries out band girdle, and band girdle width is
0.5-1.0cm, as deep as xylem.
In methods described, transplant the 5th year, proceeded as follows in the full-bloom stage of seedling：5 flowers of each inflorescence are only protected
Middle disc flower is stayed, other wholes are removed.
The genotype is that the homozygous apple varieties of R1R1 can be ' Jin Shuai ' apple, ' new Red Star ' apple, ' cold richness ' apple
Really, ' loud, high-pitched sound ' apple or ' Fuji apple ' apple.
In methods described, used as male parent, the apple variety B is used as female parent for the apple variety A.
The present invention also protects a pair of special primers pair, the single strand dna and sequence table as shown in the sequence 1 of sequence table
Single strand dna composition shown in sequence 2.The function of the special primer pair is Apple breeding.
The present invention also protects the special primer to the application in Apple breeding.
Apple new lines can be obtained using the method for the present invention, the apple new lines, in the course of cultivation without bagging,
Human cost and material cost are greatlyd save, there is great economic worth for Apple Industry, highly widelyd popularize
Using.The method provided using the present invention, cultivation integrated quality is excellent, easy coloring apple variety, and bagging is exempted to China's apple
The foundation of cultivation system, cost-saving synergistic are significant.Result shows that Apple breeding method provided by the present invention is that extremely have
Effect.The present invention has substantial worth for Apple breeding.
Brief description of the drawings
Fig. 1 is for purplish red No. 1, purplish red No. 2, purplish red No. 3, red crisp No. 1, red crisp No. 2, red crisp No. 4, red crisp No. 8 and red crisp No. 9
Fruit properties comparing.
Fig. 2 is branch, leaf, flower, fruit and the genotype identification result of purplish red No. 3.
Fig. 3 is that purplish red No. 2, red crisp No. 1, red crisp No. 2 and red crisp No. 4 fruit properties compare.
Fig. 4 is the genotype identification result and pulp extract solution photo of part individual plant in 868 plants of hybrid seedlings.
Fig. 5 is easy coloring apple new lines ' red heart 2 ' features of fruits.
Fig. 6 is easy coloring apple new lines ' red heart 31 ' features of fruits.
Fig. 7 is easy coloring apple new lines ' red heart 40 ' features of fruits.
Fig. 8 is easy coloring apple new lines ' red heart 41 ' features of fruits.
Fig. 9 is easy coloring apple new lines ' red heart 42 ' features of fruits.
Figure 10 is easy coloring apple new lines ' red heart 43 ' features of fruits.
Figure 11 is easy coloring apple new lines ' red heart 44 ' features of fruits.
Specific embodiment
Following embodiment facilitates a better understanding of the present invention, but does not limit the present invention.Experiment in following embodiments
Method, unless otherwise specified, is conventional method.Test material used in following embodiments, unless otherwise specified, is certainly
What routine biochemistry reagent shop was commercially available.Quantitative test in following examples, is respectively provided with three repetitions and tests, and as a result makes even
Average.Nutritive cube volume used in embodiment is about 350mL.Bright and clean implication is：Apple pericarp does not have scar.
' tal fibre ALMA ' apple and ' summer red meat ' apple are 2 strains in Xinjiang red meat apple (R6R1), refer to ' tal fibre
ALMA ' and ' summer red meat ' bibliography：Chen Xuesen, crystalline substance etc., Xinjiang red meat apple first generation of hybrid fruit properties heredity become
The excellent strain evaluation of different and functional form apple, Scientia Agricultura Sinica, 2014,47 (11)：2193-2204.
' Jin Shuai ' apple, ' new Red Star ' apple, ' loud, high-pitched sound ' apple and ' Fuji apple ' apple have existing apple variety, carry
And their bibliography：Chen Xuesen, pungent training just etc., the effect of marshal and Jin Shuai in Variety of Apple seed selection, Shandong agricultural
College journal, 1994,25 (2)：236—248.
' cold richness ' apple, refers to its bibliography：Li Huaiyu, Qiao Fengqi, the incubation of cool humid zone and its social effect,
Agricultural University Of Shenyang's journal, 1998,29 (l):37-40.
The preparation method of a great number of elements mother liquor of MS minimal mediums：By 1650mg NH4NO3、1900mg KNO3、440mg
CaCl2·2H2O、370mg MgSO4·7H2O and 170mg KH2PO4It is dissolved in water and is settled to 1L with water.
Seedling medium：The use that the seedling medium used in embodiments of the invention is produced for Shandong Yi Jing gardens Co., Ltd
In the plug media of flower plants and nursery stock plant, there is production and selling in general each flower plants and nursery stock company.
Single plant is referred to as strain, can obtain new lines from clip branch grafting above.
The detection of soluble solid is shown in document using conventional saccharometer method：Chen Xuesen, crystalline substance etc., Fedtsch
The first generation of hybrid fruit properties hereditary variation of meat apple and the excellent strain evaluation of functional form apple, Scientia Agricultura Sinica, 2014,47 (11)：
2193-2204。
The detection method of Anthocyanin Content is shown in document：Chen Xuesen, crystalline substance etc., Xinjiang red meat apple first generation of hybrid fruit properties
Hereditary variation and the excellent strain evaluation of functional form apple, Scientia Agricultura Sinica, 2014,47 (11)：2193-2204.
Embodiment 1, the genotype of each existing apple variety determine
Apple to be measured is ' tal fibre ALMA ' apple, ' summer red meat ' apple, ' Jin Shuai ' apple, ' new Red Star ' apple, ' cold
It is rich ' apple, ' loud, high-pitched sound ' apple or ' Fuji apple ' apple.
The genomic DNA of apple pulp to be measured is extracted, the primer pair constituted using F1 and R1 is entered performing PCR and expanded, and then presses
Following standard interpretation genotype：If pcr amplification product is a kind of and is 500bp, the genotype of apple to be measured is R6R6 homozygosis
Type；If pcr amplification product is a kind of and is 382bp, the genotype of apple to be measured is that R1R1 is homozygous；If PCR amplifications are produced
Thing is two kinds and is 500bp and 382bp, and the genotype of apple to be measured is R1R6 heterozygous.
F1 (sequence 1 of sequence table)：5’-TCAAATAGGAGGGGAATGAAG-3’；
R1 (sequence 2 of sequence table)：5’-CAGAAGCAAACACTGACAAGTT-3’.
The genotype of each apple to be measured is as follows：' tal fibre ALMA ' apple is R1R6 heterozygous；
' summer red meat ' apple is R1R6 heterozygous；
' Jin Shuai ' apple is that R1R1 is homozygous；
' new Red Star ' apple is that R1R1 is homozygous；
' cold richness ' apple is that R1R1 is homozygous；
' loud, high-pitched sound ' apple is that R1R1 is homozygous；
' Fuji apple ' apple is that R1R1 is homozygous.
Through sequencing, the sequence of the pcr amplification product of the 500bp of each apple is all consistent, such as the institute of sequence 3 of sequence table
Show.Through sequencing, the sequence of the pcr amplification product of the 382bp of each apple is all consistent, as shown in the sequence 4 of sequence table.
Embodiment 2, initiative R6R6 homozygous genotype breeding parents
First, F is created1Segregating population
(1) F is created1Segregating population -1
1st, (being carried out in Xinjiang Luntai County) is hybridized
In April, 2006 by the pollen of ' Jin Shuai ' apple to emasculation after ' tal fibre ALMA ' apple pollinate, harvest F1
Hybrid seed, by F1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning (purpose is by meeting
Chilling requirement releases seed dormancy).
2nd, nurse young plants in hothouses
In December, 2006, the seed for completing step 1 is seeded in into the nutritive cube equipped with seedling medium, and (each nutritive cube is sowed
3-5 seed) culture highly is lignifying (usually sowing 2-3 month) at 8-15cm and root neck to seedling, then will be in fact
Raw transplantation of seedlings to the new nutritive cube (each nutritive cube transplants 1 plant) equipped with seedling medium is cultivated；Culture in this step
Condition is：25 DEG C, daily illumination 12 hours (intensity of illumination 3000lx), it is every since seed is sprouted to pour within 7-10 days one time of nutrition liquid
(a great number of elements mother liquor of MS minimal mediums is diluted with water into 10 times of volumes as nutrient solution), every time each nutritive cube are poured
40-50ml。
3rd, hybrid seedling field planting and " growth promoting promotees flower contracting juvenile phase " treatment (being carried out in Taian Shandong city)
2 months 2007, every mu applied organic fertilizer and (is used in the present embodiment in the selective nursery (crop field) for planting seedling is drafted
Organic fertilizer be the cattle manure for fully becoming thoroughly decomposed) 6000kg and water heavy real；In April, 2007 completes growing directly from seeds for step 2 by 1200 plants
Seedling is colonized in selective nursery (seeding row spacing is 1m × 2m, and every mu is about colonized 333 plants), and in April, 2008 has the seed selection of seedling in field planting
Every mu applies organic fertilizer (organic fertilizer used in the present embodiment is the cattle manure for fully becoming thoroughly decomposed) 6000kg and pours in time again in garden
Water；Before applying organic fertilizer in April, 2008 from field planting seedling, foliage top dressing one is carried out with 0.3% aqueous solution of urea every two weeks
Secondary (every mu applies 5-10L every time), carries out once (every mu every time of foliage dressing with 0.4% potassium dihydrogen phosphate aqueous solution every two weeks
Apply 5-10L), foliage top dressing and foliage dressing are alternately；After applying organic fertilizer from April, 2008,0.4% phosphorus is used every two weeks
Acid dihydride aqueous solutions of potassium carries out foliage dressing once (every mu of addition 5-10L every time).
(2) F is created1Segregating population -2
1st, (being carried out in Xinjiang Luntai County) is hybridized
In April, 2006 by the pollen of ' new Red Star ' apple to emasculation after ' tal fibre ALMA ' apple pollinate, harvest
F1Hybrid seed, by F1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning (purpose is by meeting
Chilling requirement releases seed dormancy).
2nd, with the step 2 in step (1).
3rd, with the step 3 in step (1).
(3) F is created1Segregating population -3
1st, (being carried out in Xinjiang Luntai County) is hybridized
In April, 2006 by the pollen of ' cold richness ' apple to emasculation after ' tal fibre ALMA ' apple pollinate, harvest F1
Hybrid seed, by F1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning (purpose is by meeting
Chilling requirement releases seed dormancy).
2nd, with the step 2 in step (1).
3rd, with the step 3 in step (1).
(4) F is created1Segregating population -4
1st, (being carried out in Shandong Yantai City Mouping District) is hybridized
In April, 2006 by the pollen of ' tal fibre ALMA ' to emasculation after ' Fuji apple ' apple pollinate, harvest F1It is miscellaneous
Seed is handed over, by F1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning, and (purpose is by meeting need
Cold releases seed dormancy).
2nd, with the step 2 in step (1).
3rd, with the step 3 in step (1).
(5) F is created1Segregating population -5
1st, (being carried out in Shandong Yantai City Mouping District) is hybridized
In April, 2006 by the pollen of ' summer red meat ' to emasculation after ' Fuji apple ' apple pollinate, harvest F1Cenospecies
Son, by F1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning, and (purpose is by meeting chilling requirement
To release seed dormancy).
2nd, with the step 2 in step (1).
3rd, with the step 3 in step (1).
2nd, R6R6 is homozygous and R1R6 heterozygous breeding parents for initiative
Method according to embodiment 1 identifies the genotype of each plant.
, step (1), step (2), step (3), step (4) and step (5) 5 F1 segregating populations in 2009
868 plants of hybrid seedlings result successively, successively therefrom selects some plants of plant：Purplish red No. 1, purplish red No. 2, purplish red No. 3, red crisp 1
Number, red crisp No. 2, red crisp No. 4, red crisp No. 8 and red crisp No. 9.
The fruit of purplish red No. 1, purplish red No. 2, purplish red No. 3, red crisp No. 1, red crisp No. 2, red crisp No. 4, red crisp No. 8 and red crisp No. 9
Real proterties is relatively shown in Fig. 1.The branch of purplish red No. 3, leaf, flower, fruit and genotype identification result are shown in Fig. 2.Purplish red No. 2, red crisp 1
Number, red crisp No. 2 and red crisp No. 4 fruit properties compare and see Fig. 3.The genotype identification knot of part individual plant in 868 plants of hybrid seedlings
Fruit and pulp extract solution photo are shown in that (red 1, red 2, red 4, red 5, red 6, red 7 and red 8 represent the list that different pericarps are red to Fig. 4
Plant, green 1, green 2, green 3, green 4, green 5, green 6, green 7 and green 8 represent the individual plant that different pericarps are green).
The pericarp of red crisp No. 1, red crisp No. 2, red crisp No. 4, red crisp No. 8 and red crisp No. 9 is red.Purplish red No. 1, purplish red 2
Number and the pulp of purplish red No. 3 be aubergine.The branch of purplish red No. 1, purplish red No. 2 and purplish red No. 3, leaf, flower, pericarp and pulp etc. are each
Part and each stage of development are aubergine.
It is homozygous that the genotype of purplish red No. 1, purplish red No. 2 and purplish red No. 3 is R6R6.Red crisp No. 1, red crisp No. 2, red crisp 4
Number, the genotype of red crisp No. 8 and red crisp No. 9 be R1R6 heterozygous.
The genotype of purplish red No. 1 is that R6R6 is homozygous, and the color of pulp extract is aubergine.Pericarp is red individual plant
Genotype be R1R6 heterozygous, the color of pulp extract is red or pale red.Pericarp is the genotype of the individual plant of green
For R1R1 is homozygous, the color of pulp extract is colourless.
Purplish red No. 1 maturity period pulp Anthocyanin Content is 26.7U.g-1FW.Purplish red No. 2 maturity periods pulp Anthocyanin Content is
23.9U.g-1FW.Purplish red No. 3 maturity periods pulp Anthocyanin Content is 24.5U.g-1FW.Red crisp No. 1 maturity period pulp anthocyanin contains
It is 12.2U.g to measure-1FW.Red crisp No. 2 maturity periods pulp Anthocyanin Content is 2.2U.g-1FW.Red crisp No. 4 maturity periods pulp cyanine
Glycosides content is 7.5U.g-1FW.Purplish red No. 1, purplish red No. 2 and it is purplish red No. 3 show anthocyanin synthesis capability it is stronger, be easier colour
Outstanding feature, further to cultivate, integrated quality is excellent, easy coloring apple variety provides parent.
Refer to that the document of purplish red No. 1 is as follows：Wang Yan, Liu great Liang, Wang Chuanzeng, Song Yang, Chen Xiaoliu, Zhang Yanmin, Chen Xuesen,
' purplish red No. 1 ' red meat apple pulp inoxidizability and anthocyanin are analyzed, gardening journal, 2012,39 (9)：1551-1558.
Refer to that the document of purplish red No. 2 is as follows：Haifeng Xu, Wang Nan, Jiang Shenghui, Wang Yicheng, Liu Jingxuan, Qu Changzhi, Wang get Yun,
Zuo Weifang, Zhang Jing, Ji Xiaohao, Zhang Zongying, Mao Zhiquan, Chen Xuesen, the molecule mechanism of functional form apple strain flavonoids difference, in
State's agricultural sciences, 2015.
Refer to that the document of red crisp No. 1 is as follows：Haifeng Xu, Wang Nan, Jiang Shenghui, Wang Yicheng, Liu Jingxuan, Qu Changzhi, Wang get Yun,
Zuo Weifang, Zhang Jing, Ji Xiaohao, Zhang Zongying, Mao Zhiquan, Chen Xuesen, the molecule mechanism of functional form apple strain flavonoids difference, in
State's agricultural sciences, 2015.
Refer to that the document of red crisp No. 2 is as follows：Haifeng Xu, Wang Nan, Jiang Shenghui, Wang Yicheng, Liu Jingxuan, Qu Changzhi, Wang get Yun,
Zuo Weifang, Zhang Jing, Ji Xiaohao, Zhang Zongying, Mao Zhiquan, Chen Xuesen, the molecule mechanism of functional form apple strain flavonoids difference, in
State's agricultural sciences, 2015.
Refer to that the document of red crisp No. 4 is as follows：Haifeng Xu, Wang Nan, Jiang Shenghui, Wang Yicheng, Liu Jingxuan, Qu Changzhi, Wang get Yun,
Zuo Weifang, Zhang Jing, Ji Xiaohao, Zhang Zongying, Mao Zhiquan, Chen Xuesen, the molecule mechanism of functional form apple strain flavonoids difference, in
State's agricultural sciences, 2015.
Embodiment 3, cultivation easy coloring apple variety
1st, it is returned (being carried out in Shandong Yantai City Mouping District)
In April, 2010, take the pollen of purplish red No. 1, to emasculation after ' loud, high-pitched sound ' apple pollinate, harvest BC1Cenospecies
Son, by BC1Hybrid seed is put into 1-3 DEG C of fresh keeping chamber of refrigerator Stratificated treatment 60d or so after cleaning, and (purpose is by meeting chilling requirement
To release seed dormancy).
2nd, nurse young plants in hothouses (carried out in Taian Shandong)
In December, 2010, the BC that step 1 is obtained1Hybrid seed is seeded in nutritive cube (each battalion equipped with seedling medium
Support alms bowl and sow 3-5 seed) culture highly is lignifying (usually individual month of sowing 2-3) at 8-15cm and root neck to seedling,
Then transplantation of seedlings to the new nutritive cube (each nutritive cube transplants 1 plant) equipped with seedling medium that will grow directly from seeds is cultivated；This step
In condition of culture be：25 DEG C, daily illumination 12 hours (intensity of illumination 3000lx), it is every since seed is sprouted to pour one within 7-10 days
Secondary nutrient solution (a great number of elements mother liquor of MS minimal mediums is diluted with water to 10 times of volumes as nutrient solution), every time each battalion
Foster alms bowl pours 40-50ml.
3rd, hybrid seedling field planting (being carried out in Shandong Guan County)
2 months 2011, every mu applied organic fertilizer and (is used in the present embodiment in the selective nursery (crop field) for planting seedling is drafted
Be the cattle manure for fully becoming thoroughly decomposed) 6000kg and water heavy real；In April, 2011, the seedling direct planting that 3600 plants of steps 2 are obtained
In selective nursery, in April, 2011 in field planting has the selective nursery of seedling every mu apply again organic fertilizer (used in the present embodiment for
The cattle manure for fully becoming thoroughly decomposed) 6000kg and water in time.
4th, hybrid seedling band girdle treatment
In May, 2014, the trunk to each seedling at 20cm from the ground carries out band girdle, and band girdle width is 0.5-1.0cm,
As deep as xylem.The purpose of this operation is to promote the accumulation and bud differentiation of nutriment, to shorten juvenile phase, ahead of time result.
5th, flower and fruit management
The 4-5 months in 2015, each seedling full-bloom stage carry out flower thinning (5 flowers of each inflorescence only retain in disc flower, its
He all removes).Young fruit not bagging, Routine Management.
6th, easy coloring apple new plant seed selection
2015, screen easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp flower of fruit
Blue or green glycosides content is 0.010mgg-1More than FW), obtain ' red heart 2 ' and ' red heart 31 '.The fruit of ' red heart 2 ' apple is special
Levy and see Fig. 5.The features of fruits of ' red heart 31 ' apple is shown in Fig. 6.
Embodiment 4, cultivation easy coloring apple variety
Replace ' loud, high-pitched sound ' apple with ' Fuji apple ' apple, other are with embodiment 3.
2015, screen easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp flower of fruit
Blue or green glycosides content is 0.010mgg-1More than FW), obtain ' red heart 40 '.The features of fruits of ' red heart 40 ' apple is shown in Fig. 7.
Embodiment 5, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' purplish red No. 2 ' apple, other are with embodiment 3.
2015, screen easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp flower of fruit
Blue or green glycosides content is 0.010mgg-1More than FW), obtain ' red heart 41 '.The features of fruits of ' red heart 41 ' apple is shown in Fig. 8.
Embodiment 6, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' purplish red No. 2 ' apple, other are with embodiment 4.
2015, screen easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp flower of fruit
Blue or green glycosides content is 0.010mgg-1More than FW), obtain ' red heart 42 '.The features of fruits of ' red heart 42 ' apple is shown in Fig. 9.
Embodiment 7, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' purplish red No. 3 ' apple, other are with embodiment 3.
2015, from screening easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp of fruit
Anthocyanin Content is 0.010mgg-1More than FW), obtain ' red heart 43 '.The features of fruits of ' red heart 43 ' apple is shown in Figure 10.
Embodiment 8, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' purplish red No. 3 ' apple, other are with embodiment 4.
2015, screen easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp flower of fruit
Blue or green glycosides content is 0.010mgg-1More than FW), obtain ' red heart 44 '.The features of fruits of ' red heart 44 ' apple is shown in Figure 11.
Embodiment 9, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' red crisp No. 1 ' apple, other are with embodiment 3.
2015, from screening easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp of fruit
Anthocyanin Content is 0.010mgg-1More than FW), obtain ' red heart 45 '.
Embodiment 10, cultivation easy coloring apple variety
Replace ' purplish red No. 1 ' apple with ' red crisp No. 1 ' apple, other are with embodiment 4.
2015, from screening easy coloring apple new plant (screening criteria：The fruit face of fruit is bright and clean, light, the pulp of fruit
Anthocyanin Content is 0.010mgg-1More than FW), obtain ' red heart 46 '.
' loud, high-pitched sound ' Apple and ' red heart 2 ' Apple, ' red heart 31 ' Apple, ' red heart 40 ' Apples
Reality, ' red heart 41 ' Apple, ' red heart 42 ' Apple, ' red heart 43 ', ' red heart 44 ' Apple, ' red heart
No. 45 ' Apple and ' red heart 46 ' Apple be relatively shown in Table 1.
The comparing of the easy coloring apple new lines of table 1 and check variety ' loud, high-pitched sound ' fruit properties
Result shows, ' red heart 2 ', ' red heart 31 ', ' red heart 40 ', ' red heart 41 ', ' red heart 42 for newly selecting
Number ', ' red heart 43 ', ' red heart 44 ', the mean fruit weight of ' red heart 45 ' and ' red heart 46 ' in more than 100g, it is solvable
Property solid content, pericarp and pulp Anthocyanin Content be all remarkably higher than check variety ' loud, high-pitched sound ', in the case of not bagging, fruit
Real any surface finish, light.The above results show, the method provided using the present invention can be bred as that Comprehensive Traits are excellent, anthocyanin
Synthesis capability is strong, the Variety of Apple of easy coloring.
Claims (2)
1. a pair of special primers pair, the list shown in the sequence 2 of single strand dna and sequence table as shown in the sequence 1 of sequence table
Ssdna molecule is constituted.
2. special primer described in claim 1 is to the application in Apple breeding.
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| CN108377901B (en) * | 2017-02-21 | 2021-11-02 | 山东农业大学 | Continuous multi-generation bud mutation seed selection and method for continuously improving apple red character by combining with crossbreeding |
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| CN112831520B (en) * | 2021-01-07 | 2022-07-12 | 山东农业大学 | Application of R1R6 genotype apples as receptor material in preparation of red-meat transgenic apples |
| CN114868647B (en) * | 2022-06-11 | 2024-01-23 | 河北省农林科学院石家庄果树研究所 | Breeding method of fresh-eating and storage-resistant II-type red-meat apples |
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| 红肉苹果愈伤组织诱导及红色性状发育机理研究;陈学森;《山东农业大学硕士论文》;20150907;第32页第1段，第33页表2 * |
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