CN105359963A - 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
It is the key comprehensively building up well-off society that Rural Poverty Population realizes shaking off poverty, one of goal task that Ye Shi China " 13 " period is the most arduous.And economical descending, agricultural product price continues to drop, and " yellow case " policy of agriculture subsidy has also arrived " ceiling " substantially, " floor " constantly lifting of production cost; Under the Double Pressures of " ceiling " and " floor ", the sustainable high-efficient development of Apple Industry and Practices Sustainable Increasing Income of Farmers, one will expand apple consumption market, and two reduce costs and price, improves labor productivity and labor productivity, cost-saving synergistic.
Apple Bags cultivation to have the history of more than 10 year in China's large-area applications, be the key technology that China's present stage high quality apple is produced, and to raising fruit appearance quality, controls the harm of disease worm, reduces residue of pesticide and played important function.But in recent years, along with the increase year by year of labor cost, by fruit bagging, pluck the orchard recruitment cost that bag causes and become the main input cost of current apple production; Finding shows, approximately average 5000 yuan/mu of the production cost in current China apple orchard, wherein bagging, to pluck the cost that bag causes be 2600 yuan, accounts for more than 50% of total production cost; China's current apple orchard gross area 3,700 ten thousand mu, drops into bagging every year, plucks the expense of bag up to 96,200,000,000 yuan; And the soluble solid content of Apple with Fruit-bagged Treatment, total sugar content reduce, acidity improve, Studies of The Aromatic Substances content reduce, local flavor is thin out; Peeling hardness reduces, and fruit face cuticula, wax coat are thinning, respiratory metabolism and moisture loss aggravation, 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 without bagization cultivation, reduce the orchard amount of labour used and labour cost, improve fruit interior quality, become the problem that orchard worker is in the urgent need to address.Be not simple not bagging without bag cultivation, but set up modern wide row high level cadre cultivation mode by research, improve orchard ventilation and penetrating light condition, reduce damage by disease and insect occurrence index, strengthen fertilizer and drop into and reduce the series of measures such as chemical fertilizer (N is fertile) usage amount, make the apple of not bagging fruit face fineness, painted etc. in reach or substantially reach the standard of Apple with Fruit-bagged Treatment.The apple variety wherein cultivating colorability strong is one of key measure exempting from pocket culture.
Apple storage property is good, supply cycle is long, fruit contain higher proportion, human body be easier to absorb free polyphenol (flavonoids), have well anti-oxidant, antitumor, prevent cardiovascular and cerebrovascular disease and protect the effects such as liver, healthy nutritive value is high, have the good reputation of " one day apple, doctor is away from me ", considerable country is all classified as major consumers fruit and is recommended energetically in the world; Anthocyanin is the important component of flavonoids, not only has healthy nutritive value, is the constituent element of nutritional quality, and is the main matter that the Red color trait growth of mcintosh kind and exterior quality are formed.Therefore, the biosynthetic research of apple anthocyanin receives extensive concern, achieves many impressive progresses.
Summary of the invention
The object of this invention is to provide a kind of easy coloring apple variety cultivating method.
Apple breeding method provided by the invention, comprises the steps:
(1) apple variety A and apple variety B is hybridized, obtain hybrid seed; The apple variety of the homozygous or R1R6 heterozygous of described apple variety A to be genotype be R6R6; The apple variety that described apple variety B to be genotype be R1R1 is homozygous;
(2) hybrid seed sowing also nursery step (1) obtained, obtains seedling;
(3) seedling step (2) obtained is transplanted to land for growing field crops, grows after fruit do not carry out bagging to fruit until it, and screening obtains the plant with specific trait; Described specific trait is: the pulp Anthocyanin Content of fruit is 0.010mgg
-1more than FW;
Judge that the described genotypic method of apple variety is as follows:
With the genomic DNA of apple to be measured for template, the primer pair of the single strand dna shown in sequence 2 of the single strand dna shown in sequence 1 of sequence table and sequence table composition is adopted to carry out pcr amplification, then by following standard interpretation genotype: if pcr amplification product is a kind of and is 500bp, the genotype of apple to be measured is that R6R6 is homozygous; 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 amplification product is two kinds and is 500bp and 382bp, the genotype of apple to be measured is R1R6 heterozygous.
Described specific trait specifically can be: the fruit face of fruit is bright and clean, bright, and the pulp Anthocyanin Content of fruit is 0.010mgg
-1more than FW.
Described genotype is that the homozygous apple of R6R6 can be purplish red No. 1, purplish red No. 2 or purplish red No. 3.
Described genotype is that the apple of R1R6 heterozygous can be red crisp No. 1.
In described step (2), before carrying out described sowing, first described hybrid seed is carried out 1-3 DEG C of Stratificated treatment with breaking dormancy.The time of described Stratificated treatment specifically can be 60 days.
In described step (2), the condition of carrying out described nursery is: 25 DEG C, illumination every day 12 hours, intensity of illumination 3000lx, from seed germination, every 7-10 days waters one time of nutrition liquid.
In described step (2), the condition of carrying out described nursery is specially: described hybrid seed is seeded in the nutritive cube (each nutritive cube sowing 3-5 grain seed) that seedling medium is housed and is cultured to seedling height and is 8-15cm and collar place lignification (being generally sowing 2-3 month), is then transplanted by seedling to the new nutritive cube (each nutritive cube transplants 1 strain) that seedling medium is housed and cultivates; The condition of described cultivation is: 25 DEG C, illumination every day 12 hours (intensity of illumination 3000lx), and from seed germination, every 7-10 days waters one time of nutrition liquid (the macroelement mother liquor of MS minimal medium is diluted with water to 10 times of volumes and is nutrient solution), each each nutritive cube waters 40-50ml.
In described step (3), the method of carrying out cultivation management described large Tanaka is as follows: apply fertilizer 6000kg and to water depositing in water real drafting the large Tanaka every mu planting seedling, apply fertilizer 6000kg again and water in time after field planting seedling large Tanaka every mu.Described fertilizer specifically can be the cattle manure fully become thoroughly decomposed.
In described method, transplant the 4th year, carry out band girdle at overhead 20cm place to the trunk of seedling, band girdle width is 0.5-1.0cm, deeply reaches xylem.
In described method, transplant the 5th year, proceed as follows at the full-bloom stage of seedling: disc flower during 5 flowers of each inflorescence only retain, other all remove.
Described genotype is that the homozygous apple variety of R1R1 can be ' Jin Shuai ' apple, ' new Red Star ' apple, ' cold rich ' apple, ' loud, high-pitched sound ' apple or ' Fuji apple ' apple.
In described method, described apple variety A is as male parent, and described apple variety B is as female parent.
The present invention also protects a pair special primer pair, is made up of the single strand dna shown in the sequence 2 of the single strand dna shown in the sequence 1 of sequence table and sequence table.The right function of described special primer is Apple breeding.
The present invention also protects described special primer to the application in Apple breeding.
Adopt method of the present invention can obtain apple new lines, this apple new lines, in the course of cultivation without the need to bagging, greatly save human cost and material cost, for Apple Industry, there is great economic worth, highly widely popularize application.Adopt method provided by the invention, cultivate the apple variety of excellent, the easy coloring of integrated quality, the foundation of pocket culture system is exempted to China's apple, cost-saving synergistic is significant.Result shows, Apple breeding method provided by the present invention is highly effective.The present invention has substantial worth for Apple breeding.
Accompanying drawing explanation
Fig. 1 is purplish red No. 1, purplish red No. 2, purplish red No. 3, red crisp No. 1, red crisp No. 2, red crisp No. 4, the comparison of the fruit properties of red crisp No. 8 and red crisp No. 9.
Fig. 2 be purplish red No. 3 branch, leaf, flower, fruit and genotype identification result.
Fig. 3 is purplish red No. 2, red crisp No. 1, red crisp No. 2 and red crisp No. 4 fruit properties compare.
Fig. 4 is genotype identification result and the pulp extract photo of part individual plant in 868 strain hybrid seedlings.
Fig. 5 is easy coloring apple new lines ' No. 2, red heart ' features of fruits.
Fig. 6 is easy coloring apple new lines ' No. 31, red heart ' features of fruits.
Fig. 7 is easy coloring apple new lines ' No. 40, red heart ' features of fruits.
Fig. 8 is easy coloring apple new lines ' No. 41, red heart ' features of fruits.
Fig. 9 is easy coloring apple new lines ' No. 42, red heart ' features of fruits.
Figure 10 is easy coloring apple new lines ' No. 43, red heart ' features of fruits.
Figure 11 is easy coloring apple new lines ' No. 44, red heart ' features of fruits.
Embodiment
Following embodiment is convenient to understand the present invention better, but does not limit the present invention.Experimental technique in following embodiment, if no special instructions, is conventional method.Test material used in following embodiment, if no special instructions, is and purchases available from routine biochemistry reagent shop.Quantitative test in following examples, all arranges and repeats experiment for three times, results averaged.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), mention the bibliography of ' tal fibre ALMA ' and ' summer red meat ': Chen Xuesen, Zhang Jing etc., Xinjiang red meat apple first generation of hybrid fruit properties genetic variation and the excellent strain evaluation of 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 all have existing apple variety, mention their bibliography: Chen Xuesen, Xin Pei is firm, marshal and the effect of Jin Shuai in Variety of Apple seed selection, Journal of Shandong agri.Univ, 1994,25 (2): 236-248.
' cold rich ' apple, mentions its bibliography: Li Huaiyu, Qiao Fengqi, the incubation of cool humid zone and social effect thereof, Agricultural University Of Shenyang's journal, 1998,29 (l): 37-40.
The preparation method of the macroelement mother liquor of MS minimal medium: by 1650mgNH
4nO
3, 1900mgKNO
3, 440mgCaCl
22H
2o, 370mgMgSO
47H
2o and 170mgKH
2pO
4water-soluble and be settled to 1L with water.
Seedling medium: the seedling medium adopted in embodiments of the invention is the plug media of planting for flower plants and nursery stock that Yi Jing gardens, Shandong Co., Ltd produces, and all there is production and selling in general each flower plants and nursery stock company.
Single plant is called strain, from clip branch grafting above, can obtain new lines.
The detection of soluble solid adopts conventional saccharometer method, sees document: Chen Xuesen, Zhang Jing etc., Xinjiang red meat apple first generation of hybrid fruit properties genetic variation 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, Zhang Jing etc., Xinjiang red meat apple first generation of hybrid fruit properties genetic variation and the excellent strain evaluation of functional form apple, Scientia Agricultura Sinica, 2014,47 (11): 2193-2204.
The genotype of embodiment 1, each existing apple variety is determined
Apple to be measured is ' tal fibre ALMA ' apple, ' summer red meat ' apple, ' Jin Shuai ' apple, ' new Red Star ' apple, ' cold rich ' apple, ' loud, high-pitched sound ' apple or ' Fuji apple ' apple.
Extract the genomic DNA of apple pulp to be measured, adopt the primer pair of F1 and R1 composition to carry out pcr amplification, then by following standard interpretation genotype: if pcr amplification product is a kind of and is 500bp, the genotype of apple to be measured is that R6R6 is homozygous; 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 amplification product is two kinds and is 500bp and 382bp, 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 rich ' 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 order-checking, the sequence of the pcr amplification product of the 500bp of each apple is all consistent, all as shown in the sequence 3 of sequence table.Through order-checking, the sequence of the pcr amplification product of the 382bp of each apple is all consistent, all as shown in the sequence 4 of sequence table.
Embodiment 2, initiative R6R6 homozygous genotype breeding parent
One, F is created
1segregation population
(1) F is created
1segregation population-1
1, (carrying out in Luntai County, Xinjiang) is hybridized
The pollen of ' Jin Shuai ' apple is pollinated to ' tal fibre ALMA ' apple after emasculation by April, 2006, results F
1hybrid seed, by F
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, nurse young plants in hothouses
In December, 2006, the planting seed of completing steps 1 is cultured to seedling height in the nutritive cube (each nutritive cube sowing 3-5 grain seed) that seedling medium is housed and is 8-15cm and collar place lignification (being generally sowing 2-3 month), then seedling is transplanted the extremely new nutritive cube (each nutritive cube transplants 1 strain) that seedling medium is housed and cultivate; Condition of culture in this step is: 25 DEG C, illumination every day 12 hours (intensity of illumination 3000lx), and from seed germination, every 7-10 days waters one time of nutrition liquid (the macroelement mother liquor of MS minimal medium is diluted with water to 10 times of volumes and is nutrient solution), each each nutritive cube waters 40-50ml.
3, hybrid seedling field planting and " short long short flower contracting virgin phase " process (carrying out in Taian Shandong city)
In February, 2007, apply fertilizer (fertilizer adopted in the present embodiment is the cattle manure fully become thoroughly decomposed) 6000kg and to water depositing in water real drafting in the selective nursery (land for growing field crops) of planting seedling every mu; In April, 2007, in selective nursery, (seeding row spacing was 1m × 2m by the seedling direct planting of 1200 strain completing steps 2, every mu of about field planting 333 strain), there is in April, 2008 in the selective nursery of seedling every mu apply fertilizer (fertilizer adopted in the present embodiment is the cattle manure fully become thoroughly decomposed) 6000kg again and water in time in field planting; Before applying fertilizer year April from field planting seedling to 2008, within every two weeks, carry out top dressing once (every mu applies 5-10L at every turn) with 0.3% aqueous solution of urea, within every two weeks, carry out foliar application once (every mu applies 5-10L at every turn) with 0.4% potassium dihydrogen phosphate aqueous solution, top dressing and foliar application hocket; After applying fertilizer from April, 2008, within every two weeks, carry out foliar application once (every mu adds 5-10L at every turn) with 0.4% potassium dihydrogen phosphate aqueous solution.
(2) F is created
1segregation population-2
1, (carrying out in Luntai County, Xinjiang) is hybridized
The pollen of ' new Red Star ' apple is pollinated to ' tal fibre ALMA ' apple after emasculation by April, 2006, results F
1hybrid seed, by F
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, the step 2 in same step ().
3, the step 3 in same step ().
(3) F is created
1segregation population-3
1, (carrying out in Luntai County, Xinjiang) is hybridized
The pollen of ' cold rich ' apple is pollinated to ' tal fibre ALMA ' apple after emasculation by April, 2006, results F
1hybrid seed, by F
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, the step 2 in same step ().
3, the step 3 in same step ().
(4) F is created
1segregation population-4
1, (carrying out in Mouping District, Shandong Yantai City) is hybridized
The pollen of ' tal fibre ALMA ' is pollinated to ' Fuji apple ' apple after emasculation by April, 2006, results F
1hybrid seed, by F
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, the step 2 in same step ().
3, the step 3 in same step ().
(5) F is created
1segregation population-5
1, (carrying out in Mouping District, Shandong Yantai City) is hybridized
The pollen of ' summer red meat ' is pollinated to ' Fuji apple ' apple after emasculation by April, 2006, results F
1hybrid seed, by F
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, the step 2 in same step ().
3, the step 3 in same step ().
Two, the homozygous and R1R6 heterozygous breeding parent of R6R6 is formulated
The genotype of each plant is identified according to the method for embodiment 1.
2009, the 868 strain hybrid seedlings result successively of step (), step (two), step (three), step (four) and step (five) 5 F1 segregation populations, successively therefrom selects some strain plant: 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.
Purplish red No. 1, purplish red No. 2, purplish red No. 3, red crisp No. 1, red crisp No. 2, red crisp No. 4, the fruit properties of red crisp No. 8 and red crisp No. 9 relatively see Fig. 1.The branch of purplish red No. 3, leaf, flower, fruit and genotype identification the results are shown in Figure 2.Purplish red No. 2, red crisp No. 1, red crisp No. 2 and red crisp No. 4 fruit properties compare and see Fig. 3.In 868 strain hybrid seedlings, Fig. 4 (red 1, red 2, red 4, red 5, red 6, red 7 and red 8 to represent different pericarps be red individual plant, green 1, green 2, green 3, green 4, green 5, green 6, green 7 and green 8 represent different pericarps be green individual plant) is shown in by the genotype identification result of part individual plant and pulp extract photo.
Red crisp No. 1, red crisp No. 2, red crisp No. 4, the pericarp of red crisp No. 8 and red crisp No. 9 is redness.Purplish red No. 1, the pulp of purplish red No. 2 and purplish red No. 3 is aubergine.Purplish red No. 1, the branch of purplish red No. 2 and purplish red No. 3, leaf, flower, each several part such as pericarp and pulp and each developmental stage be aubergine.
Purplish red No. 1, that the genotype of purplish red No. 2 and purplish red No. 3 is R6R6 is homozygous.Red crisp No. 1, red crisp No. 2, red crisp No. 4, the genotype of red crisp No. 8 and red crisp No. 9 is R1R6 heterozygous.
The genotype of purplish red No. 1 is that R6R6 is homozygous, and the color of pulp extract is aubergine.Pericarp is the genotype of the individual plant of redness is R1R6 heterozygous, and the color of pulp extract is red or pale red.Pericarp is the genotype of the individual plant of green is that R1R1 is homozygous, and the color of pulp extract is colourless.
Purplish red No. 1 maturing stage pulp Anthocyanin Content is 26.7U.g
-1fW.Purplish red No. 2 maturing stage pulp Anthocyanin Content are 23.9U.g
-1fW.Purplish red No. 3 maturing stage pulp Anthocyanin Content are 24.5U.g
-1fW.Red crisp No. 1 maturing stage pulp Anthocyanin Content is 12.2U.g
-1fW.Red crisp No. 2 maturing stage pulp Anthocyanin Content are 2.2U.g
-1fW.Red crisp No. 4 maturing stage pulp Anthocyanin Content are 7.5U.g
-1fW.Purplish red No. 1, purplish red No. 2 and purplish red No. 3 show anthocyanin synthesis capability more by force, more easily painted outstanding feature, in order to cultivate further, integrated quality is excellent, the apple variety of easy coloring provides parent.
Mention 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 non-oxidizability and anthocyanin are analyzed, gardening journal, 2012,39 (9): 1551-1558.
Mention 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, Scientia Agricultura Sinica, 2015.
Mention 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, Scientia Agricultura Sinica, 2015.
Mention 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, Scientia Agricultura Sinica, 2015.
Mention 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, Scientia Agricultura Sinica, 2015.
Embodiment 3, cultivation easy coloring apple variety
1, backcross (carrying out in Mouping District, Shandong Yantai City)
In April, 2010, get the pollen of purplish red No. 1, ' loud, high-pitched sound ' apple after emasculation is pollinated, results BC
1hybrid seed, by BC
1hybrid seed is put into 1-3 DEG C of fridge freshness retaining room Stratificated treatment about 60d (object removes seed dormancy by meeting chilling requirement) after cleaning.
2, nurse young plants in hothouses (carrying out at Taian Shandong)
In December, 2010, the BC that step 1 is obtained
1hybrid seed is seeded in the nutritive cube (each nutritive cube sowing 3-5 grain seed) that seedling medium is housed and is cultured to seedling height and is 8-15cm and collar place lignification (being generally sowing 2-3 month), is then transplanted by seedling to the new nutritive cube (each nutritive cube transplants 1 strain) that seedling medium is housed and cultivates; Condition of culture in this step is: 25 DEG C, illumination every day 12 hours (intensity of illumination 3000lx), and from seed germination, every 7-10 days waters one time of nutrition liquid (the macroelement mother liquor of MS minimal medium is diluted with water to 10 times of volumes and is nutrient solution), each each nutritive cube waters 40-50ml.
3, hybrid seedling field planting (carrying out in Guan County, Shandong)
In February, 2011, apply fertilizer (cattle manure for fully becoming thoroughly decomposed adopted in the present embodiment) 6000kg and to water depositing in water real drafting in the selective nursery (land for growing field crops) of planting seedling every mu; In April, 2011, the seedling direct planting 3600 strain steps 2 obtained, in selective nursery, to have in the selective nursery of seedling every mu apply fertilizer (cattle manure for fully becoming thoroughly decomposed adopted in the present embodiment) 6000kg again and water in time in April, 2011 in field planting.
4, hybrid seedling band girdle process
In May, 2014, carry out band girdle at overhead 20cm place to the trunk of each seedling, band girdle width is 0.5-1.0cm, deeply reaches xylem.The object of this operation is the accumulation and the flower bud differentiation that promote nutriment, with shortening the juvenile phase, ahead of time result.
5, flower and fruit management
The 4-5 month in 2015, the full-bloom stage of each seedling carry out flower thinning (5 flowers of each inflorescence only retain in disc flower, other all remove).Young fruit is bagging not, Routine Management.
6, the planting of easy coloring apple is selected good strains in the field for seed and is educated
2015, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg to the new plant of screening easy coloring apple
-1more than FW), obtain ' No. 2, red heart ' and ' No. 31, red heart '.The features of fruits of ' No. 2, red heart ' apple is shown in Fig. 5.The features of fruits of ' No. 31, red heart ' 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, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg to the new plant of screening easy coloring apple
-1more than FW), obtain ' No. 40, red heart '.The features of fruits of ' No. 40, red heart ' 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, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg to the new plant of screening easy coloring apple
-1more than FW), obtain ' No. 41, red heart '.The features of fruits of ' No. 41, red heart ' 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, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg to the new plant of screening easy coloring apple
-1more than FW), obtain ' No. 42, red heart '.The features of fruits of ' No. 42, red heart ' 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 the new plant of screening easy coloring apple, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg
-1more than FW), obtain ' No. 43, red heart '.The features of fruits of ' No. 43, red heart ' 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, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg to the new plant of screening easy coloring apple
-1more than FW), obtain ' No. 44, red heart '.The features of fruits of ' No. 44, red heart ' 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 the new plant of screening easy coloring apple, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg
-1more than FW), obtain ' No. 45, red heart '.
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 the new plant of screening easy coloring apple, (screening criteria: the fruit face of fruit is bright and clean, bright, the pulp Anthocyanin Content of fruit was 0.010mgg
-1more than FW), obtain ' No. 46, red heart '.
' loud, high-pitched sound ' Apple compares in table 1 with ' No. 46, red heart ' Apple with ' No. 2, red heart ' Apple, ' No. 31, red heart ' Apple, ' No. 40, red heart ' Apple, ' No. 41, red heart ' Apple, ' No. 42, red heart ' Apple, ' No. 43, red heart ', ' No. 44, red heart ' Apple, ' No. 45, red heart ' Apple.
Table 1 easy coloring apple new lines compares with check variety ' loud, high-pitched sound ' fruit properties
Result shows, the average fruit weight average of ' No. 2, the red heart ', ' No. 31, red heart ', ' No. 40, the red heart ' that newly select, ' No. 41, red heart ', ' No. 42, red heart ', ' No. 43, red heart ', ' No. 44, red heart ', ' No. 45, red heart ' and ' No. 46, red heart ' is at more than 100g, soluble solid content, pericarp and pulp Anthocyanin Content are all significantly higher than check variety ' loud, high-pitched sound ', when not bagging, fruit surface is bright and clean, bright.The above results shows, Comprehensive Traits is excellent, anthocyanin synthesis capability strong, the Variety of Apple of easy coloring to utilize method provided by the invention to be bred as.
Claims (9)
1. an Apple breeding method, comprises the steps:
(1) apple variety A and apple variety B is hybridized, obtain hybrid seed; The apple variety of the homozygous or R1R6 heterozygous of described apple variety A to be genotype be R6R6; The apple variety that described apple variety B to be genotype be R1R1 is homozygous;
(2) hybrid seed sowing also nursery step (1) obtained, obtains seedling;
(3) seedling step (2) obtained is transplanted to land for growing field crops, grows after fruit do not carry out bagging to fruit until it, and screening obtains the plant with specific trait; Described specific trait is: the pulp Anthocyanin Content of fruit is 0.010mgg
-1more than FW;
Judge that the described genotypic method of apple variety is as follows:
With the genomic DNA of apple to be measured for template, the primer pair of the single strand dna shown in sequence 2 of the single strand dna shown in sequence 1 of sequence table and sequence table composition is adopted to carry out pcr amplification, then by following standard interpretation genotype: if pcr amplification product is a kind of and is 500bp, the genotype of apple to be measured is that R6R6 is homozygous; 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 amplification product is two kinds and is 500bp and 382bp, the genotype of apple to be measured is R1R6 heterozygous.
2. the method for claim 1, is characterized in that: described specific trait is: the fruit face of fruit is bright and clean, bright, and the pulp Anthocyanin Content of fruit is 0.010mgg
-1more than FW.
3. method as claimed in claim 1 or 2, is characterized in that: described genotype is the homozygous apple of R6R6 is purplish red No. 1, purplish red No. 2 or purplish red No. 3.
4. method as claimed in claim 1 or 2, is characterized in that: described genotype is the apple of R1R6 heterozygous is red crisp No. 1.
5. as the method as described in arbitrary in Claims 1-4, it is characterized in that: transplant the 4th year, carry out band girdle at overhead 20cm place to the trunk of seedling, band girdle width is 0.5-1.0cm, deeply reaches xylem.
6. as the method as described in arbitrary in claim 1 to 5, it is characterized in that: transplant the 5th year, proceed as follows at the full-bloom stage of seedling: disc flower during 5 flowers of each inflorescence only retain, other all remove.
7. as the method as described in arbitrary in claim 1 to 6, it is characterized in that: described genotype is that the homozygous apple variety of R1R1 is ' Jin Shuai ' apple, ' new Red Star ' apple, ' cold rich ' apple, ' loud, high-pitched sound ' apple or ' Fuji apple ' apple.
8. a pair special primer pair, is made up of the single strand dna shown in the sequence 2 of the single strand dna shown in the sequence 1 of sequence table and sequence table.
9. special primer described in claim 8 is to the application in Apple breeding.
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| CN106718835A (en) * | 2016-12-12 | 2017-05-31 | 山东农业大学 | Flavonoids excellent germplasm ' applications of the CSR6R6 777 ' in functional form Apple breeding high |
| WO2018107852A1 (en) * | 2016-12-12 | 2018-06-21 | 山东农业大学 | Application of high-flavonoid excellent-germplasm 'csr6r6-777' to functional apple breeding |
| US11109548B2 (en) | 2016-12-12 | 2021-09-07 | Shandong Agricultural University | Application of high flavonoid elite germplasm ‘CSR6R6-777’ in breeding for functional apple |
| CN106613921A (en) * | 2017-02-21 | 2017-05-10 | 山东农业大学 | Method for continuously improving red characters of apples by combining continuous multi-generation bud mutation selection and cross breeding |
| CN106613921B (en) * | 2017-02-21 | 2018-04-24 | 山东农业大学 | Continue more for Bud mutation and its combine the method persistently improved to apple red character with crossbreeding |
| CN108377901A (en) * | 2017-02-21 | 2018-08-10 | 山东农业大学 | Continue mostly to combine for Bud mutation and its with crossbreeding the method for persistently improveing apple red character |
| CN106937597A (en) * | 2017-04-12 | 2017-07-11 | 山东农业大学 | A kind of tissue-culturing rapid propagation method suitable for high flavonoids apple new lines |
| CN106937597B (en) * | 2017-04-12 | 2018-11-27 | 山东农业大学 | A kind of tissue-culturing rapid propagation method suitable for high flavonoids apple new lines |
| CN112831520A (en) * | 2021-01-07 | 2021-05-25 | 山东农业大学 | Application of R1R6 genotype apples as receptor material in preparation of red-meat transgenic apples |
| CN114868647A (en) * | 2022-06-11 | 2022-08-09 | 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-pulp 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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