CN108243949B - Breeding method of light and simple type temperature-sensitive genic male sterile line - Google Patents
Breeding method of light and simple type temperature-sensitive genic male sterile line Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
The invention discloses a method for breeding a thermo-sensitive genic male sterile line of light simple indica rice. The method comprises the following steps: selecting parent materials, hybridizing and selfing, fertility screening combined by multiple methods, fertility observation and matching advantage determination to obtain the flag leaf oblique downward-extending rice indica type temperature-sensitive genic male sterile line with polymerized large flag leaf angle genes. When the thermo-sensitive genic male sterile line with the obliquely downward extending sword leaves created by the method is used for producing seeds of rice, the link of manually cutting leaves in the seed production of the hybrid rice at present is saved on the premise of not reducing the competitive advantage of the prepared combined yield, the labor amount is reduced, the labor intensity is reduced, the influence of leaf cutting on the normal growth of rice plants is avoided, and the green and light seed production simplification of the hybrid rice is realized.
Description
Technical Field
The invention belongs to the field of innovation of new materials for rice heterosis utilization, and particularly relates to a breeding method of a light and simple temperature-sensitive genic male sterile line.
Background
The hybrid rice has been planted in China for nearly 40 years, and the planting area of the hybrid rice is 1600 ten thousand hm in the current year2About, more than 50% of the rice planting area plays a great role in increasing the grain yield, and gradually develops to more than 20 countries. Production of the first generation of rice hybrid seed requires artificial supplementary pollination to improve the outcrossing seed set rate. To remove pollination obstacle, the male sterile line is usually cut from the upper part 1/3-1/2 of the sword leaf. Leaf cutting not only requires high labor intensity, but also requires high operation technique to avoid damage to young and tender riceEar of grain. In addition, the wound caused by leaf cutting also has adverse effects on the normal growth of rice plants. In the course group, a indica rice local variety Anhui Hui 101 with obliquely downward extending flag leaves is found in Anhui local variety resources. 130d in the whole growth period of Anhui Hui 101, the plant height of 131cm, an included angle between the sword leaf and the rice ear of about 150 degrees, the sword leaf obliquely extends downwards, and the other leaves such as the two leaves which are inverted are obliquely lifted up like normal rice. The character is stable through field observation of multiple points (fertilizer, water of tomb) for many years.
The rice sword leaf angle refers to the included angle between the sword leaf and the rice ear in the heading period. According to the method, when the angle of the rice flag leaf is larger than 90 degrees, the rice flag leaf obliquely extends downwards, and the larger the angle of the flag leaf is, the larger the oblique extending degree of the flag leaf is. So far, genetic research on morphological characters of rice flag leaves mainly focuses on improving plant types and improving light energy utilization rate, most of the genetic research on the characters of length, width, length-width ratio and flag leaf area of the flag leaves, and the research on the angles of the rice flag leaves is less reported. Genetic analysis of the angle of the sword-like leaf from Shenfu Cheng (1983), Yangyouming et al (1990) to Dong military et al (2003) mapping the QTL controlling the angle of the sword-like leaf to WandeKai et al (2005) mapping the gene for the angle of the sword-like leafllaThe angle of the sword leaves of the materials used by researchers is less than 90 degrees, the sword leaves are obliquely lifted upwards, and the method has no practical significance for hybrid rice seed production. Therefore, the research, the discovery and the utilization of new rice germplasm resources, in particular to the oblique downward extension resources of the sword leaves with a large sword leaf angle (the sword leaf angle is more than 90 degrees), have important significance for the basic research of rice genetics, the breeding application research and the isolated cloning of related genes.
All leaves in seedling stage, tillering stage and jointing stage of Wanhui 101 are the same as those in other varieties and are lifted obliquely, but after the sword leaf pillow is higher than the poured sword leaf pillow (namely in booting stage), the sword leaf begins to stretch obliquely downwards, the angle of the sword leaf is 150 degrees, and the other leaves are still lifted obliquely. The gene with large folium xiphocae angle is introduced into the thermo-sensitive genic male sterile line, and can be freely matched with rice varieties to breed two-line hybrid rice new varieties. The hybrid rice seed production is carried out by utilizing the temperature-sensitive genic male sterile line with the oblique downward extension of the sword leaves, the link of cutting leaves of the male sterile line can be avoided, the seed production area is about 250 ten thousand mu per year in China according to the reduction of 150 yuan (1 labor day) of the labor cost per mu of the seed production field, and the seed production cost can be reduced by about 3.75 hundred million yuan. Moreover, with the development of national economy and society, a great deal of rural labor is transferred to cities, and the labor-saving, light and simple seed production technology of hybrid rice has important significance for continuously utilizing the heterosis of the rice to ensure national food safety.
Disclosure of Invention
The invention aims to provide a method for breeding a light simple indica rice thermo-sensitive genic male sterile line, the male sterile line has the genetic characteristic that the sword leaves extend obliquely downwards (the angle of the sword leaves is more than 120 degrees), other characters are similar to the thermo-sensitive genic male sterile line of the same type, and the male sterile line can be freely matched with conventional rice varieties to breed two-line hybrid rice new varieties.
The invention provides a method for breeding a thermo-sensitive genic male sterile line of light simple indica rice. The method comprises the following steps:
(1) hybridizing and selfing rice indica type temperature-sensitive genic male sterile lines which are derived from local variety resources Anhui Hui 101 (with a sword leaf angle of 150 degrees) of Anhui indica rice, Annong S (such as Guang Mi S, Guangzhan 63S and the like) and 1892S (dwarf purple S, Z913S and the like) to obtain F with different combinations2Seed generation;
(2) each combination F2Respectively planting about 3000 plants in the generation group, and screening thermo-sensitive genic male sterile single plants with the plant height of more than 120 degrees and the plant height of less than 85cm from the generation group;
(3) establishing a screening garden, selecting a single plant with the lower sterile starting point temperature (the angle of the sword leaves is more than 120 degrees) which obliquely extends downwards from the sword leaves by pollen microscopic examination and fertility observation under the environmental pressure of low temperature (23.0-23.5 ℃), selfing until the characters are stable, and recombining and polymerizing the sterile genes controlling the low starting point temperature and the genes controlling the large sword leaf angles;
(4) and through sterility identification, the indica type thermo-sensitive genic male sterile line integrating the large xiphoid leaf angle gene and the thermo-sensitive genic male sterile gene is bred.
In the breeding method of the invention, the temperature-sensitive genic male sterile line adopted in (1) is a main source of fertility genes of the indica type temperature-sensitive genic male sterile line widely applied in the current production, and is a genetic tool of two-line hybrid rice.
In the breeding method of the invention, in order to obtain the variant material in (2), a certain number of parental plants are planted as a control at the same time, and F is used as a reference2The group of sword leaf angles are divided into different grades.
In the breeding method, the hybridization of two different types of materials in the step (1), the sword leaf angle difference screening in the step (2) and the artificial environmental pressure stress in the step (3) aim to ensure that the sterile gene controlling the low starting point temperature and the large sword leaf angle gene are recombined and polymerized into a whole so as to obtain a practical sword leaf obliquely downward extending temperature-sensitive genic male sterile line.
In the breeding method of the present invention, conventional techniques can be employed for the methods not particularly pointed out. Such as observation of sterile line fertility stage sowing, identification of pollen abortion type, investigation of bagging selfing seed setting rate, emasculation by a warm soup method, artificial climate box low-temperature treatment, stubble cutting propagation and the like.
The rice obtained by the breeding method is a temperature-sensitive male sterile line with oblique downward extension of flag leaves and low sterile starting point temperature.
The temperature sensitive male nuclear sterile line is one kind of rice line with fertility controlled by recessive nuclear gene and under specific temperature condition, and has fertility conversion and sterility manifested as pollen abortion and degeneration and normal stigma development.
The fertility conversion refers to that the pollen is inactive, male sterile and incapable of selfing and fructifying at a higher temperature, and only can be fertilized and fructified by foreign pollen, and the restorer line and the matched group thereof can be utilized in production to produce hybrid seeds in a mode of artificial pollination; under the condition of lower temperature, the material shows male fertility, is self-bred and fructified, and is used for breeding in production.
The fertility transition starting point temperature refers to a temperature value at which the fertility of the temperature-sensitive genic male sterile line is transformed, and the temperature at which the fertility of the sterile line is changed from fertility to sterility at a higher temperature is referred to as sterility starting point temperature. In production, the sterile line with low sterile starting point temperature has less risk when being applied.
The restorer line is a normally-developed rice line containing dominant restorer gene and temperature-sensitive genic male sterile lineHybridization to yield F1The fertility of the generation is normal and can be self-bred and fructified.
The indica type temperature-sensitive genic male sterile line with oblique downward extending folium xiphocae obtained by the breeding method can be used for cultivating a new indica type temperature-sensitive genic male sterile line with oblique downward extending folium xiphocae; the flag leaf obliquely downward extending indica type temperature-sensitive genic male sterile line obtained by the breeding method can be used for breeding two-line hybrid rice new varieties. The thermo-sensitive genic male sterile line with oblique downward extending flag leaves and the two-line hybrid rice bred by the sterile line have normal phenotype. Experiments prove that the indica type flag leaf obliquely downward extending temperature-sensitive genic male sterile line has obvious fertility conversion, low sterile starting point temperature, longer sterile time period, complete abortion, 99.96 percent of sterility degree and 100 percent of bagging selfing failure rate; the sterile line has other characters similar to the thermo-sensitive genic sterile line of the same type except that the flag leaf extends downwards in a slant way, and can be freely matched with a conventional rice variety to breed a new two-line hybrid rice variety; the yield traits of the prepared hybrid rice combination, such as single effective ear, total grain number per ear, maturing rate, thousand grain weight and the like, are not obviously different from those of the common two-line hybrid rice.
The breeding method of the invention has the following purposes in the aspect of cultivating two-line hybrid rice:
1. the indica type temperature-sensitive genic male sterile line with oblique downward extension of the sword leaves bred by the method can be used for breeding a new rice temperature-sensitive genic male sterile line with oblique downward extension of the sword leaves;
2. the indica type temperature-sensitive genic male sterile line which is bred by utilizing the method and has obliquely downward extending sword leaves can be used for cultivating a new two-line hybrid rice variety;
3. when the temperature-sensitive genic male sterile line with the obliquely downward extending sword leaves is used for hybrid rice seed production, the link of cutting away the sword leaves of the male sterile line can be avoided, the labor intensity is reduced, the labor amount is reduced, the seed production cost is reduced, and the light and simplified seed production of the hybrid rice is realized.
The invention has the beneficial effects that:
the invention provides a method for breeding a thermo-sensitive genic male sterile line of indica rice with oblique downward extension of sword leaves by polymerizing a large sword leaf angle gene. The large flag leaf angle gene and the temperature sensitive nuclear sterile gene are firstly polymerized into a whole, so that a system capable of randomly matching and breeding a new two-line hybrid rice variety is created, the link of cutting off the flag leaf of the male sterile line in hybrid rice seed production in production is solved, the labor intensity is reduced, the labor capacity is reduced, the seed production cost is reduced, and the hybrid rice seed production is realized.
Detailed Description
The indica rice restorer line with oblique downward extending sword leaf is hybridized with the temperature-sensitive genic male sterile line material to lead the gene to be freely recombined and polymerized, the sword leaf oblique downward extending sterile material with the sword leaf angle larger than 120 degrees is screened in a separated group, and the sword leaf angle gene and the temperature-sensitive genic male sterile line are polymerized by manual selection under the environmental pressure to cultivate the temperature-sensitive genic male sterile line with oblique downward extending sword leaf, thereby achieving the aim of slightly simplifying seed production of the hybrid rice.
Example 1: sword leaf oblique downward extending temperature-sensitive genic male sterile line breeding process
1.1 materials and methods
Taking Anhui Hui 101, a local variety indica rice resource from Anhui province as a male parent, and taking 6 temperature-sensitive genic male sterile lines from Annong S (such as Mirabilis S, Guangzhan 63S and the like) and 1892S (dwarf purple S, Z913S and the like) as a female parent; sowing the parents in the Yannan Ling water base in three stages at 1, 10 and 20 months in 2011, transplanting after 28 days, and planting and row spacing of 13.3 × 16.5 cm; after the ear emergence and flowering of the male parent Anhui Hui 101, selecting a single plant which has just taken ears but does not yet flower from the temperature-sensitive genic male sterile line, castrating by a warm soup method, bagging with Anhui Hui 101 for artificial pollination respectively, and preparing hybrid seeds of different temperature-sensitive genic male sterile line combinations (Table 1).
TABLE 1 matching situation of Anhui Hui 101 and different indica type thermo-sensitive genic male sterile lines
| Combination of | Maternal origin | Source of male parent | Number of hybrid seeds |
| Mirabilis grandiflora S/Anhui 101 | Rice institute of agricultural institute of Anhui province | Rice institute of agricultural institute of Anhui province | 54 granules |
| Accounts for 63S/Anhui recovery 101 | Liaoning province farm courtyard and Fengle | Rice institute of agricultural institute of Anhui province | 38 granules |
| Xinan S/Anhui Hui 101 | Anhuquan Yinqiao (Alaska-silver-chemical family) | Rice institute of agricultural institute of Anhui province | 63 granules |
| New second S/Anhui recovery 101 | Rice institute of agricultural institute of Anhui province | Rice institute of agricultural institute of Anhui province | 45 granules |
| Shi/Wanhui 101 | Rice institute of agricultural institute of Anhui province | Rice institute of agricultural institute of Anhui province | 91 granules |
| Z913S/Anhui recovery 101 | Rice institute of agricultural institute of Anhui province | Rice institute of agricultural institute of Anhui province | 39 granules |
1.2 Breeding of thermo-sensitive genic male sterile line with oblique downward extension of sword leaf
(1) F assigned to each combination on day 4, month 27 of 20121Sowing seeds in a fertilizer-mixing base, transplanting in 5 months and 27 days, planting row spacing 13.3 multiplied by 16.5cm, and harvesting in the same combination after heading maturity to obtain F2Seeds;
(2) f combined in 2012 winter2Planting seeds in the Yannan Lingshi base, each combination of F2About 3000 plants are planted in groups, and a single plant which is obliquely extended and downwards extended from sword leaves and is segmentally fruited and possibly subjected to temperature-sensitive genic sterility is selected for seed reservation, and 6F plants with different combinations are selected23-10 selected individual plants are different in the group;
(3) sowing in late ten days of the combined fertilizer 4 months in the positive season after 2013 years, and sowing in early 12 months of the lingering water in winter, so that the combined fertilizer and Hainan alternate generation are added, and each generation preferably selects a single plant according to target characters until the strain is stable;
(4) and (3) carrying out fertility observation and identification on the selected stable lines by using stage sowing and an artificial climate box, and breeding 4 indica-type flag leaf obliquely downward-extending temperature-sensitive genic male sterile lines including 101S of Wanmian jasmine, 101S of Wanxin, 101S of Wanpurple and 913S of Wanxi in the autumn of 2017.
Example 2: fertility screening of temperature-sensitive genic male sterile line with oblique downward extension of sword leaf under natural condition
Planting different strains of the thermo-sensitive genic male sterile line with the oblique downward extension of the sword-like leaves in a seed selection garden, selecting a single spike which is just not bloomed from the spike to the full spike period, taking glume anthers of branches at the middle part of the spike for microscopic examination, taking out all 6 anthers in each glume flower during microscopic examination, putting the glume anthers on a glass slide, dripping an appropriate amount of 1% I-KI solution, fully mashing and dyeing the anthers by using tweezers, and then putting the glume anthers under a 10 multiplied by 10 microscope to observe the pollen type; selecting a similar single spike self-bred bag while sampling, and checking the self-bred fructification condition after 20 days; selecting from single plants with no pollen or a small amount of typical pollen grains according to the results of microscopic examination and bagging self-pollination fructification.
Example 3: fertility observation
In 2016, sowing starts from 4 months and 12 days to 7 months and ends at 12 days after sowing at 12 days, and in 10 stages each 10d at 1 stage, observing the fertility dynamic of the flag leaf oblique downward-extending temperature-sensitive genic male sterile line in each stage. Planting about 100 plants per period, the row spacing of the plants is 13.3 multiplied by 16.5cm, and the seedling age is 28 days; from the beginning of heading (day 13/7) to the end of heading, 5 ears are taken out every 1-2 days for microscopic examination of pollen, and the pollen abortion degree is observed; meanwhile, selecting a similar 10-spike bagging, and investigating the selfing and fruiting rate when the seedlings are mature. The results show that: microscopic examination between 7 month, 13 days and 9 months, 8 days, pollen abortion rate of 99.95% -100%, bagging self-sterile rate of 0, microscopic examination between 9 months, 12 days and 23 days, pollen abortion rate of 68.91% -96.82%, bagging self-sterile rate of 13.75% -43.82%, which indicates that the observed Jianye obliquely downward-extending temperature-sensitive genic male-sterile line is longer in fertile sterility period and obvious in sterile fertile conversion period (Table 2).
Table 22016 years fertility observation of flag leaf obliquely downward extending temperature-sensitive genic male sterile line under natural condition of fertilizer combination
Example 4: determination of grouping advantages of flag leaf obliquely downward-extending temperature-sensitive genic male sterile line
(1) 4 flag leaf oblique downward-extending thermo-sensitive genic male sterile lines: 101S of Wan jasmine, 101S of Wanxin, 101S of Wanzi and 913S of Wan; 7 restorer lines: anxiang No. 2, Anxiang No. 15, R998, R727, Zhongxian WH26, Huazhan, Yanggao No. 6;
(2) planting 4 flag leaf obliquely downward extending temperature-sensitive genic male sterile lines and 7 restorers in a cemetery base in 2016 (12) years, performing warm soup emasculation by using the 4 flag leaf obliquely downward extending temperature-sensitive genic male sterile lines as female parents in the heading and flowering period, and bagging and pollinating the 7 restorers respectively to obtain 28 hybrid combinations;
(3) 28 hybrid combinations and 2 controls are sowed in a compost base at 5 month and 8 days in 2017, the hybrid combinations and the controls are transplanted at 12 months in 6 months, the plant-row spacing is 16.5 multiplied by 26.5cm, the hybrid combinations are planted singly, the yield and the characters are investigated in the mature period, and the results are listed in table 3.
Investigation of combined yield traits of the combined thermo-sensitive genic male sterile lines with the oblique downward extension of the sword leaves in Table 34
| Combination of | Plant height/cm | Effective ear of single plant | Total number of grains per ear | Fruit set/percent | Thousand grain weight/g |
| Wan Mo No. 101S/Anxiang No. 2 | 125.2 | 11.5 | 236.4 | 82.5 | 26.6 |
| Wan Mo 101S/Anxiang 15 | 132.4 | 9.8 | 217.2 | 86.3 | 25.7 |
| Wan Mo 101S/R998 | 131.9 | 11.9 | 212.3 | 80.5 | 27.3 |
| Wan jasmine 101S/R727 | 128.1 | 12.6 | 207.5 | 88.1 | 27.1 |
| Wan Mo 101S/Zhongnong WH26 | 127.6 | 10.3 | 226.4 | 83.4 | 27.5 |
| 101S/Huazhan of Wan jasmine | 126.5 | 10.8 | 229.9 | 82.3 | 25.6 |
| No. 101S/Yanggao No. 6 Wan jasmine | 132.3 | 12.7 | 238.1 | 85.1 | 25.2 |
| Anhui New No. 101S/Anxiang No. 2 | 126.8 | 11.0 | 213.4 | 86.7 | 27.6 |
| Anhui New 101S/Anxiang 15 | 126.1 | 12.4 | 245.2 | 88.4 | 26.8 |
| Anhui New 101S/R998 | 127.5 | 10.3 | 239.1 | 83.2 | 26.6 |
| Anhui New 101S/R727 | 126.2 | 10.0 | 217.3 | 84.1 | 27.6 |
| Anhui Xin 101S/Zhongxian WH26 | 119.7 | 9.2 | 226.1 | 80.9 | 27.9 |
| Anhui New 101S/Huazhan | 128.2 | 9.1 | 257.1 | 85.6 | 28.5 |
| Anhui New No. 101S/Yanggao No. 6 | 128.6 | 10.5 | 233.8 | 81.5 | 27.8 |
| Wan Zi 101S/Anxiang No. 2 | 118.3 | 9.8 | 250.2 | 87.3 | 28.3 |
| Wan Zi 101S/Anxiang 15 | 119.2 | 8.7 | 247.6 | 82.8 | 28.7 |
| Wan Zi 101S/R998 | 118.5 | 10.3 | 236.4 | 84.1 | 27.9 |
| Wan Zi 101S/R727 | 120.1 | 11.7 | 216.4 | 82.9 | 28.1 |
| Wan Zi 101S/Zhongnong WH26 | 121.5 | 12.1 | 213.8 | 84.6 | 27.8 |
| Wan Zi 101S/Huazhan | 115.7 | 11.5 | 237.7 | 85.2 | 28.2 |
| Wan Zi 101S/Yanggao No. 6 | 116.5 | 11.0 | 226.2 | 86.7 | 27.6 |
| Anhui 913S/Anxiang No. 2 | 122.6 | 13.2 | 232.0 | 87.2 | 28.1 |
| Wan 913S/Anxiang 15 | 128.3 | 10.5 | 208.7 | 85.4 | 28.3 |
| Anhui 913S/R998 | 124.3 | 11.9 | 222.5 | 83.2 | 28.3 |
| Anhui 913S/R727 | 129.7 | 10.5 | 243.2 | 85.6 | 28.1 |
| Wan 913S/Zhongxian WH26 | 131.4 | 9.9 | 211.6 | 81.8 | 28.5 |
| Wan 913S/Huazhan | 128.4 | 9.3 | 220.9 | 86.1 | 28.4 |
| Anhui 913S/Yanggao No. 6 | 123.6 | 12.1 | 206.3 | 84.3 | 28.9 |
| Average | 125.2 | 10.9 | 227.6 | 84.5 | 27.6 |
| Yangliangyou No. 6 (CK 1) | 127.1 | 10.5 | 227.8 | 86.1 | 27.4 |
| II Yo 838 (CK 2) | 125.2 | 11.1 | 215.2 | 87.3 | 28.1 |
(4) As shown in Table 3, the effective ears of the 28 combinations are 10.9 per plant, between 8.7 and 13.2 and between 2 controls; the average total grain number per spike is 227.6, which is basically consistent with No. 6 Yangliangyou and is higher than II you 838; the average setting percentage was 84.5%, which was slightly lower than the two controls; the average thousand kernel weight was 27.6g, which was between 2 controls. Therefore, the yield and the character of the hybrid rice variety matched by the flag leaf obliquely downward extending temperature-sensitive genic male sterile line are normal.
Claims (1)
1. The method for breeding the light and simple temperature-sensitive genic male sterile line is characterized by comprising the following steps:
(1) hybridizing and selfing indica rice resource Anhui Hui 101 with oblique downward extending xifolium and 150-degree angle with rice indica type temperature-sensitive genic male sterile line from Annong S and 1892S to obtain various combinations F2Seed generation;
(2) each combination F2Planting about 3000 plants in a colony, and screening thermo-sensitive genic male sterile plants with oblique and downward extending xiphoid leaves and an angle of more than 120 degrees from the population;
(3) establishing a screening garden, screening single plants with low sterile starting point temperature, plant height less than 85cm and inclined sword leaf downward extension and angle more than 120 degrees through pollen microscopic examination and fertility observation under the environmental pressure of 23.0-23.5 ℃, selfing until the characters are stable, and recombining and polymerizing the sterile gene controlling the low starting point temperature and the sword leaf angle gene;
(4) after fertility identification, a large-flag-leaf-angle indica type temperature-sensitive genic male sterile line is bred, wherein the flag-leaf obliquely downward-extending gene and the temperature-sensitive genic male sterile gene are polymerized in the same line, and the sterile line is a light and simple temperature-sensitive genic male sterile line;
the angle of the sword leaves of the light and simple temperature-sensitive genic male sterile line is more than 120 degrees, and the plant height is less than 85 cm.
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| CN105075848A (en) * | 2015-08-07 | 2015-11-25 | 安徽省农业科学院水稻研究所 | Hybrid rice breeding method |
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| CN1218615A (en) * | 1998-11-05 | 1999-06-09 | 四川省农业科学院作物研究所 | Production method of hybrid rice |
| CN105075848A (en) * | 2015-08-07 | 2015-11-25 | 安徽省农业科学院水稻研究所 | Hybrid rice breeding method |
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