CN108935073B - Seed production method of purplish red marked three-line hybrid cotton - Google Patents
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Abstract
The invention discloses a seed production method of purple red marked three-line hybrid cotton. The purplish red marker character is transferred to the upland cotton cytoplasm male insect-resistant sterile line, the restorer line is hybridized with the upland cotton cytoplasm male insect-resistant sterile line to produce the insect-resistant three-line hybrid with the purplish red marker character, the purity can be identified in advance in the cotyledon period, compared with the traditional method that the purity of the three-line hybrid is identified by comprehensive character identification in the cotton growth period, the identification period is greatly shortened, the purity identification problem of the three-line hybrid in production is better solved, and the three-line hybrid plays a better role in production.
Description
Technical Field
The invention relates to the field of crop breeding, in particular to a seed production method of purplish red marked three-line hybrid cotton.
Background
In the world, more than 30 crops, such as corn and three-line hybrid rice, have achieved great success in the application of the hybrid vigor. Human research on cotton heterosis has been in the past for over a hundred years. According to the report of American Cotton production research institute corpus, the yield Heterosis (HP) range of the ginned cotton is 4-33%, and the average value is 15-27%; the maximum heterosis for the mcolon values was 4%, the down length was 9%, and the strength was 6.6%. The research on the heterosis of cotton is highly valued by various major cotton-producing countries, and the research on hybrid cotton is developed in China from the sixties of the last century, so that a large number of hybrid cotton varieties are bred and popularized and applied on production in a large scale, such as 29-medium cotton, 15-Shandong cotton, Handan miscellaneous 98-1, Handan miscellaneous 301 and the like.
At present, cotton hybrid production mainly adopts methods of inter-species, two-line and three-line seed production, and the inter-species seed production needs manual castration, is labor-consuming and time-consuming, has high cost and small scale, and leads to difficult guarantee of purity due to incomplete castration; the two-line method needs to eliminate 50 percent of fertile plants in the field, is easy to cause seedling shortage and ridge breaking, and has low seed production yield and low purity; the three-line method saves the links of artificial emasculation, fertile plant elimination and the like, and has the advantages of less labor, low cost and high purity. Therefore, the seed production by the three-line method is the most effective way for utilizing the heterosis of cotton. (the "cotton three-line" refers to male sterile line, maintainer line and restoring line, the male sterile line refers to variety and line with male sterile property, it is called sterile line for short, the male organ of sterile line can not normally develop, it has no pollen or can not form pollen with normal function, but the pistil is normal, it can accept external pollen and fertilize, so it can be used as female parent in producing seed, it can not emasculate, the maintainer line is used for pollinating the sterile line, the variety or line keeping its sterility, it is called maintainer line for short, the cell nucleus is identical with that of sterile line, it is only different in cytoplasm, so the maintainer line can not only keep sterility of sterile line for one generation, but also can keep its hereditary property generation, the sterile line used in producing seed can obtain seed by pollinating the maintainer line, the produced conventional cotton variety can be used as maintainer line, the source is wide; the restoring line is a variety or strain with restoring gene, after pollination, the obtained hybrid seed is restored to normal male fertility, so that it is called male sterile restoring line, called restoring line for short, and when the restoring line is used as male parent to make hybridization with sterile line to obtain hybrid seed, its progeny is fertile. )
In the aspect of cotton three-line utilization, a Hakini cotton cytoplasmic male sterile line is bred in 60 s of 20 th century abroad, the three-line matching is completed in 1973, and through years of research, a dominant combination cannot be bred for production so far. In China, Handan City agricultural science academy has been engaged in the cotton three-line application research for a long time, upland cotton cytoplasm male sterile line '104-7A' different from Haknessi cotton cytoplasm is bred in 1983, three-line matching is completed, the first medium-maturing monovalent insect-resistant three-line hybrid cotton Handan hybrid 98-1 in 2005 in China is bred, the gap of transgenic insect-resistant three-line hybrid cotton in China is filled up through examination in Hebei province and national examination in 2006, and a new approach for hybrid cotton breeding is opened up.
At present, the parents for producing the three-line hybrid seeds and the leaves of the three-line hybrid seeds are all of the conventional green leaf type, the purity of the hybrid seeds can be visually distinguished without any appearance marking characters, and the phenomenon that the cotton hybrid seeds are adulterated to be fake and are excellent frequently occurs in the market under the drive of benefits, so that the rights and interests of breeding workers cannot be guaranteed, and meanwhile, irreparable loss is brought to seed operation enterprises and cotton farmers.
Disclosure of Invention
The invention aims to provide a seed production method of purple red marked three-line hybrid cotton, which can remove impurity plants and identify the purity of the obtained three-line hybrid cotton according to purple red marked characters of plants in the early stage, and also achieves the aims of high yield, high quality and disease and insect resistance, thereby protecting the benefits of breeding workers, enterprises and cotton farmers.
The invention provides upland cotton (Gossypium hirsutum) Handan 6208 application of mauve A in breeding hybrid cotton.
The invention also provides a breeding method of hybrid cotton, which comprises the following steps: using upland cotton (Gossypium hirsutum) Handan 6208 purple red A is used as female parent, and the conventional restoring line is used as male parent to make hybridization so as to obtain the invented hybrid cotton.
Said is male parent Handan R251. The male parent may also be Handan R174 or 18R.
Any one of the hybrid cotton has a purplish red marker character; the character of having the mauve marker is shown in that the color of the cotyledon is mauve after the cotyledon is unearthed.
Any of the above hybrid cotton has insect resistance. The insect resistance can be specifically the resistance of lepidoptera insects such as cotton bollworms, pink bollworms and the like.
Any of the above-described hybrid cotton has wilt resistance and verticillium wilt resistance.
Upland cotton (Gossypium hirsutum) Handan 6208 mauve A was deposited in the general microbiological culture collection center of China Committee for culture Collection of microorganisms (CGMCC; address: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; and E, postcode: 100101), preservation number of CGMCC NO. 15381.
The purplish red marker character is transferred to the upland cotton cytoplasm male insect-resistant sterile line, the restorer line is hybridized with the upland cotton cytoplasm male insect-resistant sterile line to produce the insect-resistant three-line hybrid with the purplish red marker character, the purity can be identified in advance in the cotyledon period, compared with the traditional method that the purity of the three-line hybrid is identified through comprehensive character identification in the later growth period of cotton, the identification period is greatly shortened, the purity identification problem of the three-line hybrid in production is better solved, and the three-line hybrid plays a better role in production.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified. The quantitative tests in the following examples, all set up three replicates and the results averaged.
Purple red marker trait 27005 line: the germplasm resource material 27005 with the shape of the purple leaf marker is introduced from the research institute of cotton of Chinese academy of agricultural sciences; the public can be obtained from Handan City agricultural scientific college.
Insect-resistant sterile line Handan 6208A: reference documents: the breeding process and the cultivation technical point of a new high-yield and high-quality cotton variety Handan 6208, wherein [ J ] Chinese cotton, 2014 and 14(10) is 87.109; the public can be obtained from Handan City agricultural scientific college. Handan 6208A is a Handan 6208 sterile line, which was transformed by Handan 6208. Handan 6208 was examined in 2007 by the province of north river: ji trial cotton 2007011.
Insect-resistant sterile line Handan 750A: reference documents: the breeding and cultivation technology of transgenic insect-resistant three-line hybrid cotton Handan hybrid 301 [ J ]. Chinese cotton, 2010,37(11): 30-30%; the public can be obtained from Handan City agricultural scientific college. Handan 750A is the female parent of Handan Hete 301, Handan Hete 301 passes the national approval in 2009 with approval numbers: 2009015.
insect-resistant sterile line SGK 321A: reference documents: breeding a new variety Handan hybrid 429 of a new transgenic triple hybrid cotton variety Maweijun, Rev. people, 2008,35(2): 26-26.; the public can be obtained from Handan City agricultural scientific college. SGK321A is the female parent of Handan heter 429, Handan heter 429 was approved by the north-river province in 2006, and was approved by the state in 2007, with the approval numbers: ji dynasty cotton 2006010 and Country dynasty cotton 2007012.
Handan R251: reference documents: the breeding and cultivation technology of transgenic insect-resistant three-line hybrid cotton Handan hybrid 301 [ J ]. Chinese cotton, 2010,37(11): 30-30%; the public can be obtained from Handan City agricultural scientific college. Handan R251 is a male parent of a three-line hybrid Handan hybrid 429 and Handan hybrid 301, Handan hybrid 429 is approved by Hebei province in 2006, and is approved by the state in 2007, and the approval numbers are: ji dynasty cotton 2006010 and Country dynasty cotton 2007012.
Handan hetero 301: reference documents: the breeding and cultivation technology of transgenic insect-resistant three-line hybrid cotton Handan hybrid 301 [ J ]. Chinese cotton, 2010,37(11): 30-30%; the public can be obtained from Handan City agricultural scientific college. The product is approved by the nation in 2009 with the approval serial number: 2009015.
handan R174: reference documents: high-yield, high-quality and disease and insect resistant three-line hybrid cotton Handan hybrid 98-1 breeding [ J ] Hebei agricultural science, 2007,11(4): 88-88%; the public can be obtained from Handan City agricultural scientific college. Handan R174 is a male parent of Handan hybrid 98-1 of the three-line hybrid, Handan hybrid 98-1 is approved by Hebei province in 2005, and is approved by the state in 2006, and the approval numbers are: ji dynasty cotton 2005001 and Guo dynasty cotton 2006007.
18R reference: zho\28952, Guo hong Yuan, Zhang Jiang, etc. construction of BAC library of upland cotton male sterility restorer line 18R [ J ]. Cotton school report, 2009,21(3): 252-; the public can be obtained from Handan City agricultural scientific college. 18R is a male parent of a three-line hybrid strain sGKz8, and the sGKz8 passes the national approval in 2005, and the approval serial number is as follows: national cotton 2005001.
In the examples, "having a purple-red marker trait" is expressed in that the color of the cotyledon is purple-red after the cotyledon emerges.
Example 1 acquisition of sterile line with purplish-Red marker trait
Breeding of sterile line with purple red marker character
1. In a trial field of agricultural scientific institute of Handan, 7 months in 2006, a purplish red marked character 27005 is taken as a male parent, an insect-resistant sterile line Handan 6208A is taken as a female parent for hybridization, the purplish red marked character is transferred into an insect-resistant sterile line Handan 6208A, the purplish red marked character 27005 is taken as a recurrent parent in 2007 and 2009, the purplish red marked character is subjected to hybridization transfer and screening for more than 6 times in Hainan and Handan agricultural scientific institute trial fields, and the purplish cotton cytoplasmic male anti-insect sterile line Handan 6208 purplish red A with the purplish red marked character and optimal performance is transferred to more than 6 generations in 2009.
2. In a 7-month Handan agricultural scientific institute experimental field in 2006, a magenta-labeled character 27005 line is taken as a male parent, an insect-resistant sterile line Handan 750A is taken as a female parent, the magenta-labeled character is transferred to an insect-resistant sterile line Handan 750A, the magenta-labeled character 27005 line is taken as a recurrent parent 2007-plus 2009, and the hybrid transfer and screening are carried out for more than 6 times in Hainan and Handan agricultural scientific institute experimental fields in 2009, and the wild cotton cytoplasmic male anti-insect sterile line Handan 750 magenta A with the magenta-labeled character and optimal performance is transferred to 6 generations in 2009.
3. In a trial field of agricultural scientific institute of Handan, 7 months in 2006, a magenta-labeled character 27005 is taken as a male parent, an insect-resistant sterile line SGK321A is taken as a female parent for hybridization, the magenta-labeled character is transferred into an insect-resistant sterile line SGK321A, the magenta-labeled character 27005 is taken as a recurrent parent in 2007 and 2009 to carry out hybridization transfer and screening for more than 6 times in Hainan and Handan agricultural scientific institute trial field, and the upland cotton cytoplasm male anti-insect sterile line SGK321 magenta A with the magenta-labeled character and optimal performance is transferred to more than 6 generations in 2009.
Second, survey of sterility rate, sterility degree, insect resistance and disease resistance of sterile line
The sterile line to be tested: upland cotton cytoplasmic male anti-insect sterile line Handan 6208 mauve A, upland cotton cytoplasmic male anti-insect sterile line Handan 750 mauve A and upland cotton cytoplasmic male anti-insect sterile line SGK321 mauve A.
The method for investigating and evaluating the sterility rate and the sterility degree comprises the following steps: selecting 100 sterile cotton plants at the flowering stage of 10-20 days in 7 months, and investigating flowers which bloom on each plant at 9-10 am every day, wherein the standard is as follows: the anther of each flower on each plant is shrunken and the pollen-free flower is twisted by hand to be completely sterile, the sterile rate and the sterile degree are 100 percent, otherwise, the sterile rate and the sterile degree are not 100 percent.
② the insect resistance identification method: the daily concentration of the cotton seedling is 4000mg in 3-5 true leaf period and 25-27 month in 5-5-1The kanamycin sulfate solution is smeared on cotton leaves, the color of the smeared leaves is observed after four days, the normal leaves are insect-resistant plants, and the yellow leaves are non-insect-resistant plants, so that the insect resistance of the cotton leaves is identified.
The insect resistance is the resistance of lepidoptera pests such as cotton bollworms, pink bollworms and the like.
③ the identification method of the blight and verticillium wilt resistance: disease conditions were investigated 1 time at the peak of blight and verticillium wilt, using 5-grade disease grading standard. The disease grading criteria are as follows:
i standard for grading disease conditions of blight
Level 0: no pathological condition is observed on the exterior.
Level 1: the disease of the plant leaves is less than 25%, and the plant type is normal.
And 2, stage: 25-50% of leaves show symptoms, and the plant type is microscopically dwarfed.
And 3, level: more than 50% of leaves show symptoms, and the plant type is dwarfed.
4, level: the diseased plants wither and die.
II standard for grading etiopathic verticillium disease
Level 0: the cotton plants are healthy, have no diseased leaves and grow normally.
Level 1: less than one third of the leaves of cotton plants show symptoms.
And 2, stage: more than one third and less than two thirds of the leaves of the cotton plant show symptoms.
And 3, level: more than two thirds of the leaves of cotton plants showed symptoms and were not withered.
4, level: the cotton plants died.
The disease percentage (%) × (total number of diseased plants ÷ total number of investigated plants) × 100%.
Disease means ═ 100 [ disease strains at each level were multiplied by the sum of the corresponding progression ÷ (total strains investigated × highest progression) ].
The identification results are as follows:
firstly, 100 plants of each of three sterile lines are investigated in the cotton flowering phase in 7, 10-20 days in 2010, the sterile rate of the three sterile lines is 100%, and the anthers of the plants in the flowering phase are shrunken and have no pollen, and the sterility degree is also 100%.
② the concentration of applying 4000mg to each 100 sterile lines in 5 months and 25 days in 2010-1The kanamycin sulfate solution shows that the cotton leaves are normal and have no color change, which indicates that the three sterile lines resist cotton bollworm.
③ investigating 100 sterile lines in 2010 at 25 days in 6 months (blast disease incidence peak period) and 25 days in 8 months (verticillium wilt incidence peak period), wherein the blast disease values of Handan 6208 mauve A and Handan 750 mauve A, SGK321 mauve A are respectively 5.1, 7.6 and 9.7 through grading calculation, and the three sterile lines resist blast disease; handan 6208 mauve a and Handan 750 mauve A, SGK321 mauve a have verticillium wilt values of 20.1, 29.3 and 25.0 respectively, and Handan 6208 mauve a, Handan 750 mauve a and SGK321 mauve a belong to the type of verticillium wilt resistance.
Among the three sterile lines, Handan 6208 mauve a is superior to Handan 750 mauve a and SGK321 mauve a in blast and verticillium wilt resistance.
In conclusion, the sterility rate and sterility degree of Handan 6208 purplish red A are both 100%, and the sterile line is resistant to blight, verticillium wilt and cotton bollworm and belongs to the optimal sterile line with purplish red marker traits.
Deposit of three, Handan 6208 purple-red A
Handan 6208 purple red A is named as upland cotton (Gossypium hirsutum) Handan 6208 mauve A, which was deposited in China general microbiological culture Collection center (CGMCC; address: western road No.1, north west city of township, beijing, institute of microbiology, china academy of sciences; and E, postcode: 100101), preservation number of CGMCC NO. 15381. Upland cotton (Gossypium hirsutum) Handan 6208 magenta a is abbreviated Handan 6208 magenta a.
Example 2 selection of three lines of hybrid Cotton with purple Red marker trait
The following 3 hybridization combinations were set:
1. in 2011, Handan 6208 purple red A with purple red marker character is used as a female parent, and a conventional restorer line Handan R251 is used as a male parent for hybridization to obtain the insect-resistant three-line hybrid with purple red marker character.
2. In 2011, Handan 6208 purple red A with purple red marker character is used as a female parent, and a conventional restorer line Handan R174 is used as a male parent for hybridization to obtain the insect-resistant three-line hybrid with purple red marker character.
3. In 2011, Handan 6208 purple red A with purple red marker character is used as a female parent, and a conventional restorer line 18R is used as a male parent for hybridization to obtain the insect-resistant three-line hybrid with purple red marker character.
The assembled hybrid combinations were subjected to the self-set yield ratio test (using Handan hybrid 301 as a control) for 2012-2016 continuously for many years, wherein the average seed, lint and pre-frost lint yield results and fiber quality results for 2014-2016 for three years are shown in Table 1. The yield indices (pre-frost flower rate, clothes point, yield) in table 1 are obtained from post-harvest yield in crop breeding; the testing method of the fiber quality index (fiber length, specific strength and micronaire value) is tested by a cotton quality supervision inspection testing center of Ministry of agriculture; the method for resisting insect and blight and verticillium wilt is the same as that of example 1.
TABLE 12014-2016 average yield, quality and disease and insect resistance results in three years
The above results show that: the combination (Handan 6208 mauve A × Handan R251) is superior to the combinations (Handan 6208 mauve A × Handan R174 and Handan 6208 mauve A × 18R) in yield, quality, disease resistance and the like, and the combination has better performance, and the same female parent is best in combination comprehensive traits with Handan R251 as a male parent.
The invention provides a sterile line with purplish red mark character, which can produce three-line hybrid seeds with purplish red mark character, so that the obtained three-line hybrid seeds can be immediately subjected to impurity removal and purity identification according to the purplish red mark character of plants after cotton seedlings emerge.
Claims (9)
1. Upland cotton (Gossypium hirsutum) Handan 6208 the application of mauve A in cultivating hybrid cotton; the upland cotton (Gossypium hirsutum) Handan 6208 deposit number for mauve a: CGMCC NO. 15381.
2. The use of claim 1, wherein: the hybrid cotton has a purplish red marking character; the character of having the mauve marker is shown in that the color of the cotyledon is mauve after the cotyledon is unearthed.
3. Use according to claim 1 or 2, characterized in that: the hybrid cotton has insect resistance.
4. Use according to claim 1 or 2, characterized in that: the hybrid cotton has wilt resistance and verticillium resistance.
5. A method for breeding hybrid cotton comprises the following steps: the method of using the upland cotton (C) of claim 1Gossypium hirsutum) Handan 6208 purple red A is used as female parent, and the conventional restoring line is used as male parent to make hybridization so as to obtain the invented hybrid cotton.
6. The method of claim 5, wherein: said is male parent Handan R251.
7. The method of claim 5 or 6, wherein: the hybrid cotton has a purplish red marking character; the character of having the mauve marker is shown in that the color of the cotyledon is mauve after the cotyledon is unearthed.
8. The method of claim 5 or 6, wherein: the hybrid cotton has insect resistance.
9. The method of claim 5 or 6, wherein: the hybrid cotton has wilt resistance and verticillium resistance.
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| CN101491209B (en) * | 2009-02-25 | 2011-08-24 | 广西大学 | Perennial crossover seed-production method of annual cotton |
| CN102440180A (en) * | 2011-10-11 | 2012-05-09 | 邯郸市农业科学院 | Method for hybridizing and breeding cotton male sterility restoring line |
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| CN102499067A (en) * | 2011-11-21 | 2012-06-20 | 山东农兴种业有限责任公司 | Colored hybrid cotton with aubergine leaves and method for identifying purity and trueness of cotton |
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