CA2748306A1 - Breeding method for brassica napus l. self-incompatibility two-line hybrid - Google Patents
Breeding method for brassica napus l. self-incompatibility two-line hybrid Download PDFInfo
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Abstract
Provided is a breeding method for Brassica napus L. self-incompatibility two-line hybrid, including a breeding and propagating method for self-incompatible lines and self-incompatibility restorer lines, and a breeding method for self-incompatibility two-line hybrid and its seed production method.
Fig. 1 selected as the drawing accompanying the abstract.
Fig. 1 selected as the drawing accompanying the abstract.
Description
Title: Breeding Method for Brassica Napus L.
Self-incompatibility Two-line Hybrid Technical Field The invention relates to a breeding method for Brassica Napus L., specifically, relates to a breeding method for Brassica napus L.
self-incompatibility two-line hybrid.
Background Art A large number of researches in China and abroad indicate that the inter-varietal hybrid of Brassica Napus L. exhibits a great deal of heterosis.
Currently, the ways of utilizing heterosis of Brassica Napus L. include cytoplasmic male sterility (CMS), genic male sterility (GMS), self-incompatibility (SI) and chemical hybridization. Genic male sterility (GMS) of Brassica Napus L. has an advantage of stable sterility, however its fertility is likely to segregate, when producing seeds, 50% of the fertile plants in the sterility line needs to be pulled out at first flowering stage of Brassica Napus L.; that will reduce the yield of seed production and increase the cost of seed production, further, since it is generally difficult to completely pull out the fertile plants, that will affect the purity of seeds and increase production risk. The way of chemical hybridization has such shortages as environment pollution and low purity of the hybrid. At present, Pol cms is the well-known most valuable cytoplasmic male sterility line of Brassica Napus L. in the world;
however, its fertility is apt to be affected by environmental conditions, and the breeding processes of maintainer line and restorer line are complicated and have long periods, and have potential risk of negative effects of cytoplasma.
Comparing with other ways of utilizing heterosis of Brassica Napus L., self-incompatibility hybrid of Brassica Napus L. has advantages including short breeding period, being easy to screen a restorer line, no negative effects of cytoplasma and high yield of seed production. Some possible ways of utilizing self-incompatible lines of Brassica napus L. to prepare hybrids have been proposed previously, such as self-incompatibility single-cross hybrid (self-incompatible line A x self-incompatible line B), self-incompatibility three-line hybrid ((self-incompatible line x maintainer line) x restorer line), self-incompatibility three-way cross hybrid ((self-incompatible line A x self-incompatible line B ) x restorer line) and self-incompatibility double cross hybrid ((self-incompatible line A x self-incompatible line B) x (self-incompatible line C x self-incompatible line D)) (Breeding and utilization of rapeseed hybrid, page 142-145, editor in chief, FU Tingdong;
Hubei Science and Technology Press, August 2000). Although these ways have been proposed for many years, due to the limit of breeding method for Brassica napus L. self-incompatibility hybrid, currently the breeders have not bred any Brassica napus L. self-incompatibility hybrid which is suitable for large scale popularization in agricultural production. Aiming at the drawbacks of existing breeding methods for Brassica napus L.
self-incompatibility hybrid, the present invention provides a breeding method for Brassica napus L. self-incompatibility two-line hybrid. The method is expected to breed Brassica napus L. self-incompatibility hybrids which are suitable for large scale popularization in agricultural production, thus promoting heterosis utilization of Brassica Napus L.
Contents of the Invention The object of the present invention is to overcome drawbacks of the prior art, and provide a breeding method for Brassica napus L.
self-incompatibility two-line hybrid. The invention is a breakthrough and innovation based on the existing three-line breeding method for rapeseed. The technical solution of the invention generally includes breeding and propagation of Brassica napus L. self-incompatible lines, breeding and propagation of Brassica napus L. self-incompatibility restorer lines, breeding of Brassica napus L. self-incompatibility two-line hybrid and method of seed production thereof.
The invention will promote application of the Brassica napus L.
self-incompatibility two-line system in current heterosis breeding.
The invention is carried out through the following technical solutions:
A breeding method for Brassica napus L. self-incompatibility two-line hybrid, comprising the following steps:
Self-incompatibility Two-line Hybrid Technical Field The invention relates to a breeding method for Brassica Napus L., specifically, relates to a breeding method for Brassica napus L.
self-incompatibility two-line hybrid.
Background Art A large number of researches in China and abroad indicate that the inter-varietal hybrid of Brassica Napus L. exhibits a great deal of heterosis.
Currently, the ways of utilizing heterosis of Brassica Napus L. include cytoplasmic male sterility (CMS), genic male sterility (GMS), self-incompatibility (SI) and chemical hybridization. Genic male sterility (GMS) of Brassica Napus L. has an advantage of stable sterility, however its fertility is likely to segregate, when producing seeds, 50% of the fertile plants in the sterility line needs to be pulled out at first flowering stage of Brassica Napus L.; that will reduce the yield of seed production and increase the cost of seed production, further, since it is generally difficult to completely pull out the fertile plants, that will affect the purity of seeds and increase production risk. The way of chemical hybridization has such shortages as environment pollution and low purity of the hybrid. At present, Pol cms is the well-known most valuable cytoplasmic male sterility line of Brassica Napus L. in the world;
however, its fertility is apt to be affected by environmental conditions, and the breeding processes of maintainer line and restorer line are complicated and have long periods, and have potential risk of negative effects of cytoplasma.
Comparing with other ways of utilizing heterosis of Brassica Napus L., self-incompatibility hybrid of Brassica Napus L. has advantages including short breeding period, being easy to screen a restorer line, no negative effects of cytoplasma and high yield of seed production. Some possible ways of utilizing self-incompatible lines of Brassica napus L. to prepare hybrids have been proposed previously, such as self-incompatibility single-cross hybrid (self-incompatible line A x self-incompatible line B), self-incompatibility three-line hybrid ((self-incompatible line x maintainer line) x restorer line), self-incompatibility three-way cross hybrid ((self-incompatible line A x self-incompatible line B ) x restorer line) and self-incompatibility double cross hybrid ((self-incompatible line A x self-incompatible line B) x (self-incompatible line C x self-incompatible line D)) (Breeding and utilization of rapeseed hybrid, page 142-145, editor in chief, FU Tingdong;
Hubei Science and Technology Press, August 2000). Although these ways have been proposed for many years, due to the limit of breeding method for Brassica napus L. self-incompatibility hybrid, currently the breeders have not bred any Brassica napus L. self-incompatibility hybrid which is suitable for large scale popularization in agricultural production. Aiming at the drawbacks of existing breeding methods for Brassica napus L.
self-incompatibility hybrid, the present invention provides a breeding method for Brassica napus L. self-incompatibility two-line hybrid. The method is expected to breed Brassica napus L. self-incompatibility hybrids which are suitable for large scale popularization in agricultural production, thus promoting heterosis utilization of Brassica Napus L.
Contents of the Invention The object of the present invention is to overcome drawbacks of the prior art, and provide a breeding method for Brassica napus L.
self-incompatibility two-line hybrid. The invention is a breakthrough and innovation based on the existing three-line breeding method for rapeseed. The technical solution of the invention generally includes breeding and propagation of Brassica napus L. self-incompatible lines, breeding and propagation of Brassica napus L. self-incompatibility restorer lines, breeding of Brassica napus L. self-incompatibility two-line hybrid and method of seed production thereof.
The invention will promote application of the Brassica napus L.
self-incompatibility two-line system in current heterosis breeding.
The invention is carried out through the following technical solutions:
A breeding method for Brassica napus L. self-incompatibility two-line hybrid, comprising the following steps:
2 (1) breeding of Brassica napus L. self-incompatible line: Brassica napus L. self-incompatible line S-1300 as female parent (see: Breeding of Brassica napus L. double low (low erucic acid, low glucosinolates) self-incompatible lines, MA Chaozhi, et al, Journal of Huazhong Agricultural University, 1998 , 17 (3): 211-213) was crossed with Brassica napus L.
self-compatible line Hua Shuang No. 3 (also referred to as Huayou Shuang No. 3, they are actually the same variety; the rapeseed variety was bred by Huazhong Agricultural University in Wuhan, Hubei province of China, which was approved by Hubei Provincial Crop Variety Approval Committee on March 26, 1998, certificate number: Hubei seed certificate No. 156; it was a new variety of Brassica napus L. which has been openly popularized) to obtain F1 seeds; the self-compatibility of F1 plants were measured by compatible index method, and the self-incompatible F1 plants were subjected to bud-pollination to obtain F2, the self-compatibility of F2 plants were measured, and the self-incompatible F2 plants were selected and subjected to selfing by artificial bud-pollination to obtain F3 seeds; F3 seeds were sowed and the F3 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-pollination to obtain seeds; F4 seeds from F3 family lines in which the compatible index of individual plant is less than 1 were sowed and the F4 family lines were selected, all individual plants of the selected lines were selfed in bags and
self-compatible line Hua Shuang No. 3 (also referred to as Huayou Shuang No. 3, they are actually the same variety; the rapeseed variety was bred by Huazhong Agricultural University in Wuhan, Hubei province of China, which was approved by Hubei Provincial Crop Variety Approval Committee on March 26, 1998, certificate number: Hubei seed certificate No. 156; it was a new variety of Brassica napus L. which has been openly popularized) to obtain F1 seeds; the self-compatibility of F1 plants were measured by compatible index method, and the self-incompatible F1 plants were subjected to bud-pollination to obtain F2, the self-compatibility of F2 plants were measured, and the self-incompatible F2 plants were selected and subjected to selfing by artificial bud-pollination to obtain F3 seeds; F3 seeds were sowed and the F3 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-pollination to obtain seeds; F4 seeds from F3 family lines in which the compatible index of individual plant is less than 1 were sowed and the F4 family lines were selected, all individual plants of the selected lines were selfed in bags and
3-5 individual plants selected from every line were subjected to selfing by artificial bud-pollination to obtain F5 seeds, the F5s from F4 family lines with compatible index of less than 1 were selected as the desired Brassica napus L. self-incompatible lines; the F3 family lines with unstable self-incompatibility were repeatedly subjected to the above steps until a Brassica napus L. self-incompatible line with stable self-incompatibility was selected; the applicant thus obtained Brassica napus L.
self-incompatible line 07-P63-6 YU 34 through the above steps;
(2) breeding of Brassica napus L. self-incompatibility restorer lines:
Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatible rapeseed line Zheyou 18 (bought from China Zhejiang Academy of Agricultural Sciences), Huyou 17 (bought from China Shanghai Academy of Agricultural Sciences) and Hui 5900 (the term "Hui"
means restorer) (see: Genetic Diversity of Parents for Hybrid Breeding in Brassica napus L. Detected by RAPDs and RFLPs, MA Chaozhi, et al., Acta Agronomica Sinica, 2003, 29 (5): 701-707) to obtain F1 seeds; F1 seeds were sowed and the self-compatibility of F1 was investigated; said self-compatible rapeseed lines were sowed at the same time, and their yield and yield-related traits were investigated, the breeding materials with good field performance, wherein the compatible index of F1 combination from that breeding material and said Brassica napus L. self-incompatible line S-1300 was over 10, were selected as target restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900;
(3) breeding of Brassica napus L. self-incompatibility two-line hybrid:
Brassica napus L. self-incompatible line S-1300 and its improved self-incompatible line 07-P63-6 YU 34 as female parents were crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to breed two-line hybrid; F1 combinations and the contrast varieties were sowed in field at the same time according to randomized block design, and agronomic traits, yield, disease resistance, growth period and quality traits of the plants were investigated to select the hybrid combination with strong heterosis for subsequent use;
self-incompatible line 07-P63-6 YU 34 through the above steps;
(2) breeding of Brassica napus L. self-incompatibility restorer lines:
Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatible rapeseed line Zheyou 18 (bought from China Zhejiang Academy of Agricultural Sciences), Huyou 17 (bought from China Shanghai Academy of Agricultural Sciences) and Hui 5900 (the term "Hui"
means restorer) (see: Genetic Diversity of Parents for Hybrid Breeding in Brassica napus L. Detected by RAPDs and RFLPs, MA Chaozhi, et al., Acta Agronomica Sinica, 2003, 29 (5): 701-707) to obtain F1 seeds; F1 seeds were sowed and the self-compatibility of F1 was investigated; said self-compatible rapeseed lines were sowed at the same time, and their yield and yield-related traits were investigated, the breeding materials with good field performance, wherein the compatible index of F1 combination from that breeding material and said Brassica napus L. self-incompatible line S-1300 was over 10, were selected as target restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900;
(3) breeding of Brassica napus L. self-incompatibility two-line hybrid:
Brassica napus L. self-incompatible line S-1300 and its improved self-incompatible line 07-P63-6 YU 34 as female parents were crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to breed two-line hybrid; F1 combinations and the contrast varieties were sowed in field at the same time according to randomized block design, and agronomic traits, yield, disease resistance, growth period and quality traits of the plants were investigated to select the hybrid combination with strong heterosis for subsequent use;
(4) propagating of Brassica napus L. self-incompatible lines: the means of early sowing, low density cultivation and high fertilizer were used to cultivate strong seedlings of Brassica napus L. self-incompatible lines, 3%
NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination, the compatibility index of each plant was investigated and the seeds of the plant with compatibility index of less than 1 were harvested as breeder seeds of Brassica napus L.
self-incompatible line, the seeds were kept in refrigerator at -20 C for subsequent use; a small amount of breeder seeds were sowed in the next sowing season, 3% NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination to propagate the self-incompatible lines, and the original seeds of Brassica napus L. self-incompatible lines were obtained;
NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination, the compatibility index of each plant was investigated and the seeds of the plant with compatibility index of less than 1 were harvested as breeder seeds of Brassica napus L.
self-incompatible line, the seeds were kept in refrigerator at -20 C for subsequent use; a small amount of breeder seeds were sowed in the next sowing season, 3% NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination to propagate the self-incompatible lines, and the original seeds of Brassica napus L. self-incompatible lines were obtained;
(5) propagating of Brassica napus L. self-incompatibility restorer lines:
Brassica napus L. cytoplasmic male sterility, genic male sterility, or chemical hybridization were carried out to propagate the Brassica napus L.
self-incompatibility restorer lines (see: Breeding and utilization of rapeseed hybrid, pages 145-150, editor in chief, FU Tingdong; Hubei Science and Technology Press, Wuhan, China, 1995);
Brassica napus L. cytoplasmic male sterility, genic male sterility, or chemical hybridization were carried out to propagate the Brassica napus L.
self-incompatibility restorer lines (see: Breeding and utilization of rapeseed hybrid, pages 145-150, editor in chief, FU Tingdong; Hubei Science and Technology Press, Wuhan, China, 1995);
(6) producing the seeds of Brassica napus L. self-incompatibility two-line hybrid: Brassica napus L. self-incompatible line obtained from step (1) was served as female parent, Brassica napus L. self-incompatibility restorer line obtained from step (2) was served as male parent, wherein the female parent and male parent plants were planted under artificial isolated condition or in natural isolated area in a row ratio of 2:1 or 3:1 or 4:1, in blossoming of the Brassica napus L. plants, honeybees were used to supplement pollination, and at the end of final flowering stage of Brassica napus L. the plants of male parent rows were removed, the female parent row plants were harvested at mature stage of Brassica napus L. to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
The advantages of the invention are as follows:
1. The breeding of Brassica napus L. self-incompatibility two-line hybrid solves the technical problem that Brassica napus L.
self-incompatibility hybrid was not able to popularize and cultivate in a great area in agricultural production.
2. Compared with the existing breeding method for Brassica napus L.
three-line hybrid, only female parent and male parent are needed for the Brassica napus L. self-incompatibility two-line hybrid, maintainer line and temporary maintainer line are not involved, thus the invention has the advantages of simplified breeding process, short breeding period and high breeding efficiency.
3. Compared with the existing breeding method for Brassica napus L.
three-line hybrid, the method for propagation of female parent (Brassica napus L. self-incompatible lines) is simple.
4. Breeding method for Brassica napus L. self-incompatibility restorer lines (male parent) is simple. Since the vast majority of conventional varieties /lines of Brassica napus L. can restore the self-incompatibility of Brassica napus L. self-incompatible lines, thus readily selecting an excellent hybrid combination.
5. The yield of seed production for Brassica napus L.
self-incompatibility two-line hybrid is high and the cost of seed production is low.
For more detailed technical solutions, see "Specific Mode for Carrying out the Invention".
Description of Drawings Figure 1 is the technical road map according to the invention.
Figure 2 shows Brassica napus L. self-incompatible line S-1300 after selfing in bags and the seeding states of its hybrid F1 in the present examples, figures a-d indicates that Brassica napus L. self-incompatible line S-1300 after selfing in bag hardly produces seeds; figure e indicates the seeding state of F1 hybrid developed by crossing Brassica napus L. self-incompatible line S-1300 as female parent and conventional Brassica napus L. line Zheyou 18 as male parent after selfing in bags, which normally produces seeds.
Figure 3 is the propagation effect of Brassica napus L.
self-incompatible line S-1300 under isolation shelter condition.
Figure 4 is the production state of the seed of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Zheyou 18 as male parent with female parent and male parent in a row ratio of 2:1) under isolation shelter condition.
Figure 5 is the production state of the seed of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Zhongshuang No. 10 (an excellent Brassica napus L. variety bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, Wuhan in Hubei province, China, which had been widely popularized in agricultural production) as male parent with female parent and male parent in a row ratio of 2:1 under condition of natural isolated area, Figure 5 shows the seedling stage of Brassica napus L.
Figure 6 is the production state of the seeds of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Brassica napus L. variety Zhongshuang No. 10 as male parent with female parent and male parent in a row ratio of 2:1) under condition of natural isolated area, Figure 6 shows the pod mature stage of Brassica napus L.
Specific Mode for Carrying out the Invention In terms of the biological materials involved in the following examples, it should be understood according to the part of "Contents of the Invention"
that the applicant has made a statement to the disclosed way of obtaining the biological materials used in examples 1-2 for breeding (also referred to as crossing) (see: Certificate of Compliance 1-2). Owing to the limitation of space, said contents of Certificate of Compliance are omitted in examples 1-2, however, it should not be construed as lacking the description of the biological material in the invention.
Example 1 1. Breeding of Brassica napus L. self-incompatible lines, which comprises the following steps:
(1) Brassica napus L. self-incompatible line S-1300 as female parent was crossed with Brassica napus L. self-compatible line Hua Shuang No. 3 to obtain seeds of the first generation of hybrid (F1).
(2) The F1 seeds obtained from step (1) were sowed. The main
The advantages of the invention are as follows:
1. The breeding of Brassica napus L. self-incompatibility two-line hybrid solves the technical problem that Brassica napus L.
self-incompatibility hybrid was not able to popularize and cultivate in a great area in agricultural production.
2. Compared with the existing breeding method for Brassica napus L.
three-line hybrid, only female parent and male parent are needed for the Brassica napus L. self-incompatibility two-line hybrid, maintainer line and temporary maintainer line are not involved, thus the invention has the advantages of simplified breeding process, short breeding period and high breeding efficiency.
3. Compared with the existing breeding method for Brassica napus L.
three-line hybrid, the method for propagation of female parent (Brassica napus L. self-incompatible lines) is simple.
4. Breeding method for Brassica napus L. self-incompatibility restorer lines (male parent) is simple. Since the vast majority of conventional varieties /lines of Brassica napus L. can restore the self-incompatibility of Brassica napus L. self-incompatible lines, thus readily selecting an excellent hybrid combination.
5. The yield of seed production for Brassica napus L.
self-incompatibility two-line hybrid is high and the cost of seed production is low.
For more detailed technical solutions, see "Specific Mode for Carrying out the Invention".
Description of Drawings Figure 1 is the technical road map according to the invention.
Figure 2 shows Brassica napus L. self-incompatible line S-1300 after selfing in bags and the seeding states of its hybrid F1 in the present examples, figures a-d indicates that Brassica napus L. self-incompatible line S-1300 after selfing in bag hardly produces seeds; figure e indicates the seeding state of F1 hybrid developed by crossing Brassica napus L. self-incompatible line S-1300 as female parent and conventional Brassica napus L. line Zheyou 18 as male parent after selfing in bags, which normally produces seeds.
Figure 3 is the propagation effect of Brassica napus L.
self-incompatible line S-1300 under isolation shelter condition.
Figure 4 is the production state of the seed of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Zheyou 18 as male parent with female parent and male parent in a row ratio of 2:1) under isolation shelter condition.
Figure 5 is the production state of the seed of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Zhongshuang No. 10 (an excellent Brassica napus L. variety bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, Wuhan in Hubei province, China, which had been widely popularized in agricultural production) as male parent with female parent and male parent in a row ratio of 2:1 under condition of natural isolated area, Figure 5 shows the seedling stage of Brassica napus L.
Figure 6 is the production state of the seeds of Brassica napus L.
self-incompatibility two-line hybrid (Brassica napus L. self-incompatible line S-1300 as female parent and Brassica napus L. variety Zhongshuang No. 10 as male parent with female parent and male parent in a row ratio of 2:1) under condition of natural isolated area, Figure 6 shows the pod mature stage of Brassica napus L.
Specific Mode for Carrying out the Invention In terms of the biological materials involved in the following examples, it should be understood according to the part of "Contents of the Invention"
that the applicant has made a statement to the disclosed way of obtaining the biological materials used in examples 1-2 for breeding (also referred to as crossing) (see: Certificate of Compliance 1-2). Owing to the limitation of space, said contents of Certificate of Compliance are omitted in examples 1-2, however, it should not be construed as lacking the description of the biological material in the invention.
Example 1 1. Breeding of Brassica napus L. self-incompatible lines, which comprises the following steps:
(1) Brassica napus L. self-incompatible line S-1300 as female parent was crossed with Brassica napus L. self-compatible line Hua Shuang No. 3 to obtain seeds of the first generation of hybrid (F1).
(2) The F1 seeds obtained from step (1) were sowed. The main
7 inflorescences of F1 plants were subjected to selfing in bags during the flowering period of Brassica napus L. and the self-compatibility of F1 was estimated. If F1 exhibits self-compatibility, then F1 was subjected to selfing in bags; if F1 exhibits self-incompatibility, then F1 was subjected to selfing by bud-pollination; the F2 seeds were obtained;
(3) The F2 seeds obtained from step (2) were sowed. Each F2 plant was selfed in bags during the flowering period of Brassica napus L. The self-incompatible plants were selected and selfed by artificially bud-pollination. The seeds of each F2 plants selfed by bud-pollination were harvested.
(4) The F2 seeds of selfed by bud-pollination from step (3) with compatibility index of less than 1 were sowed to obtain F3 family line. F3 family line was selected depending on the breeding objectives such as growth situation in seedling period and growth period. All the individual plants of the selected F3 family line were selfed in bags, and each plant with excellent self-incompatibility was selected to be selfed by artificially bud-pollination.
The seeds from F3 family line selfed by bud-pollination with compatibility index of less than 1, were taken as the seeds of F4 family line during mature stage.
(5) The seeds of F4 family line were sowed. F4 family line was selected depending on breeding objectives such as growth situation at the seedling stage and growth period. All individual plants of the selected F4 family line were selfed in bags and the individual plants with excellent self-incompatibility were selected to be selfed by bud-pollination with hand.
All the F4 family lines, whose compatibility index of individual plant is less than 1, were namely the new Brassica napus L. self-incompatible lines.
(6) If there was no F4 family line whose compatibility index of individual plant is less than 1, the step (4) was repeated until a new Brassica napus L. self-incompatible line with stable self-incompatibilty was selected.
Through the above steps, the applicant has obtained Brassica napus L.
self-incompatible line 07-P63-6 YU 34.
(3) The F2 seeds obtained from step (2) were sowed. Each F2 plant was selfed in bags during the flowering period of Brassica napus L. The self-incompatible plants were selected and selfed by artificially bud-pollination. The seeds of each F2 plants selfed by bud-pollination were harvested.
(4) The F2 seeds of selfed by bud-pollination from step (3) with compatibility index of less than 1 were sowed to obtain F3 family line. F3 family line was selected depending on the breeding objectives such as growth situation in seedling period and growth period. All the individual plants of the selected F3 family line were selfed in bags, and each plant with excellent self-incompatibility was selected to be selfed by artificially bud-pollination.
The seeds from F3 family line selfed by bud-pollination with compatibility index of less than 1, were taken as the seeds of F4 family line during mature stage.
(5) The seeds of F4 family line were sowed. F4 family line was selected depending on breeding objectives such as growth situation at the seedling stage and growth period. All individual plants of the selected F4 family line were selfed in bags and the individual plants with excellent self-incompatibility were selected to be selfed by bud-pollination with hand.
All the F4 family lines, whose compatibility index of individual plant is less than 1, were namely the new Brassica napus L. self-incompatible lines.
(6) If there was no F4 family line whose compatibility index of individual plant is less than 1, the step (4) was repeated until a new Brassica napus L. self-incompatible line with stable self-incompatibilty was selected.
Through the above steps, the applicant has obtained Brassica napus L.
self-incompatible line 07-P63-6 YU 34.
8 2. Breeding of Brassica napus L. self-incompatibility restorer lines, which comprises the following steps:
(1) Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatibility Brassica napus L. varieties Zheyou 18, Huyou 17 and Hui 5900 to obtain seeds of the first generation of hybrid (F1).
(2) The F1 seeds from step (1) were sowed. All the F1 plants were selfed in bags during the flowering period of Brassica napus L. The compatibility index of F1 plants was evaluated during the mature stage.
(3) Self-compatible Brassica napus L. lines were planted in the field at the same time when the step (2) was carried out, and their yield and yield-related traits were investigated.
(4) Combining the results of steps (2) and (3), the breeding materials with good field performance were selected, namely the Brassica napus L.
self-incompatibility restorer lines, wherein the compatibility index of F1 combination obtained by crossing the breeding material and said Brassica napus L. self-incompatible line S-1300 is over 10. Through the above steps the applicant has obtained Brassica napus L. self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900.
3. Breeding of Brassica napus L. self-incompatibility two-line hybrid, which comprises the following steps:
(1) Brassica napus L. self-incompatible lines S-1300 and 07-P63-6 YU
34 were respectively as female parents crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to obtain F1 hybrid combination.
(2) According to randomized block design, F1 combinations and the contrast varieties were sowed in test field at the same time, with 2-3 replications, 3-5 rows in one area, row length of 1.9m, row width of 25cm and spacing in the plants of 17cm. During the mature stage of Brassica napus L., 10-15 plants were selected randomly from each area and the other plants in each area were harvested. Agronomic traits including plant height,
(1) Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatibility Brassica napus L. varieties Zheyou 18, Huyou 17 and Hui 5900 to obtain seeds of the first generation of hybrid (F1).
(2) The F1 seeds from step (1) were sowed. All the F1 plants were selfed in bags during the flowering period of Brassica napus L. The compatibility index of F1 plants was evaluated during the mature stage.
(3) Self-compatible Brassica napus L. lines were planted in the field at the same time when the step (2) was carried out, and their yield and yield-related traits were investigated.
(4) Combining the results of steps (2) and (3), the breeding materials with good field performance were selected, namely the Brassica napus L.
self-incompatibility restorer lines, wherein the compatibility index of F1 combination obtained by crossing the breeding material and said Brassica napus L. self-incompatible line S-1300 is over 10. Through the above steps the applicant has obtained Brassica napus L. self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900.
3. Breeding of Brassica napus L. self-incompatibility two-line hybrid, which comprises the following steps:
(1) Brassica napus L. self-incompatible lines S-1300 and 07-P63-6 YU
34 were respectively as female parents crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to obtain F1 hybrid combination.
(2) According to randomized block design, F1 combinations and the contrast varieties were sowed in test field at the same time, with 2-3 replications, 3-5 rows in one area, row length of 1.9m, row width of 25cm and spacing in the plants of 17cm. During the mature stage of Brassica napus L., 10-15 plants were selected randomly from each area and the other plants in each area were harvested. Agronomic traits including plant height,
9 branching part and yield per plant, disease resistance, growth period and yield in each area for each combination were investigated based on the breeding objectives, and the quality traits of the harvested seeds including oil content and erucic acid content were analyzed.
(3) The F1 combinations which met the breeding objectives were namely the strong heterosis combinations available for the next experiment. Through the above steps, the strong heterosis combinations according to the invention increased the production by 8-12% than that of the contrast variety Zhongyouza No. 2 (an excellent rapeseed variety bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, approved and popularized widely over China).
4. Propagating of Brassica napus L. self-incompatible lines, which comprises the following steps:
(1) Brassica napus L. self-incompatible line S-1300 was sowed every year from later August to early September under isolated conditions with spacing in the rows of 2 m and spacing between plants of I m in winter rapeseed-planting areas along the middle and lower reaches of Yangtze river, China, and Brassica napus L. self-incompatible line with large plant shape and branches as many as possible was bred. Each plant of Brassica napus L.
self-incompatible line S-1300 was encased with a 60-mesh nylon breeze tent at bolting stage, and honeybee pollination was deployed in the tent. The main inflorescences of the self-incompatible line were selfed in bags, and self-compatibility of the plant of the self-incompatible line was identified.
(2) During flowering period, the plants of Brassica napus L.
self-incompatible line S-1300 under isolated condition were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total.
(3) At mature stage of Brassica napus L. self-incompatible line S-1300, seeds of the self-incompatible line of individual plant were harvested. The compatibility index was calculated by the formula: compatibility index =
seed number / flower number, and the plants whose compatibility index is over 2 were weeded out. Contents of erucic acid and glucosinolates of the seeds of each plant were analyzed, and the seeds with erucic acid content of less than 1% and glucosinolates content of less than 30umol/g were selected as breeder seeds of Brassica napus L. self-incompatible line, and the seeds were kept in refrigerator at -20 C for subsequent use.
(4) The breeder seeds obtained from step (3) were continuously sowed in the next propagation season. In flowering period, plants of the self-incompatible line were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total. Honeybee pollination was deployed, and original seeds of Brassica napus L. self-incompatible line were harvested for use in agricultural production.
5. Propagation of Brassica napus L. self-incompatibility restorer lines:
Propagation methods of restorer lines by utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, or genic male sterility, or chemical hybridization were used to propagate the Brassica napus L.
self-incompatibility restorer lines (see: Breeding and utilization of rapeseed hybrid, page 145-150, editor in chief, FU Tingdong; Hubei Science and Technology Press, 1995) .
6. Seed production method for Brassica napus L. self-incompatibility two-line hybrid:
(1) Brassica napus L. self-incompatible line as female parent was crossed with Brassica napus L. self-incompatibility restorer line as male parent, wherein the female parent and male parent plants were planted under artificial isolated shelter or in natural isolated area in a row ratio of 2:1 or 3:1 or 4:1. The field management is the same as that of the local large field crop production.
(2) In blossoming of the plants of Brassica napus L., honeybees were arranged to assist pollination in artificial isolated shelter or natural isolated area.
(3) At the end of final flowering stage of Brassica napus L., the plants of male parent rows were removed.
(4) At mature stage of Brassica napus L., the female parent row plants were harvested to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
Example 2 (comparative test) The detail steps are as follows:
1. Using compatibility index to evaluate the self-compatibility of Brassica napus L.
At primary flowering stage, the main branches of the investigated Brassica napus L. plants were encased in bags. When there were 3-5 flowers blossoming at a main branch, all the blossoming flowers at the main branch were removed and some little flower buds at the top of inflorescence center were thinned out;
then the main branch and two side branches were encased in a sulfate paper bag and the bag was clipped tightly with paper clips. The paper bag was pulled upward every two days, while patting it to shed the pollen to assistant pollination.
If there were flowers or flower buds revealing out of the bag when the bags were pulled up, the flowers or flower buds should be removed immediately to avoid harvesting impure seeds. When the pollination of almost all flowers in bags was finished, the bags were removed and the buds and un-pollinated flowers were stricken off, a preliminary investigation of self-compatibility was carried out based on the development of rapeseed pods. After harvesting, the seeds were threshed and the compatibility index of each plant was investigated. The compatibility index of a plant was calculated by the formula: compatibility index (Self-compatibility Index, SCI) = seed number /flower number.
2. Breeding of Brassica napus L. self-incompatible line 07-P63-6 YU 34 (1) In spring of 1998, the self-incompatible line S-1300 as female parent was crossed with Hua Shuang No. 3, a semi-winter type Brassica napus L.
line in China, to obtain seeds of the first generation of hybrid (F1), in a rapeseed testing field of Huazhong Agricultural University, Wuhan in Hubei province, China.
(2) In autumn of 1999, F1 seeds were sowed (2 rows, 10 plants /row). In spring of 2000, the F1 plants were selfed in bags in blossoming to evaluate the self-compatibility of F1, and the F1 plants were selfed by artificially bud-pollination. F1 plants were harvested in early May, and the selfed F1 plants hardly went to seed.
(3) In autumn of 2000, the seeds of the selfed F1 plants were sowed to obtain F2 plants (Field number: 01-9-701, 2 rows /area, 10 plants /row, 20 F2 plants in total), in rapeseed testing field of Huazhong Agricultural University, Wuhan; in spring of 2001, 8-10 individual plants of the Brassica napus L. were selected to be selfed and selfed by bud-pollination in blossoming; when the rapeseed pods were mature in early May, the seeds of each selfed F2 plant were harvested separately, and the compatibility index was investigated, the seeds of the plant whose compatibility index is less than 1 were kept.
(4) In 2006, the Brassica napus L. self-incompatible line with stable self-incompatibility and double-low quality was obtained through the breeding method of steps (3)-(6) according to the invention.
In autumn of 2006, the seeds harvested from an individual plant with low compatibility index, low erucic acid content and low glucosinolates content, and more seeds obtained by bud-pollination, were sowed in testing field in Huazhong Agricultural University, Wuhan, and the testing number is YU 34; the plants were transplanted in an isolated shelter No. P63-6 in early November, and the field number is 07-P63-6 YU 34 , the plants were transplanted in 5 rows and has a number of about 50 in total; in spring of 2007, the shelter was encased with nylon breeze tent to isolate the foreign pollen; all the individual plants grew normally, the number of seeds by selfing was small, compatibility index was less than 1, the content of erucic acid <1%, and the content of glucosinolates <30umol/g, they met the double-low standard.
3. Breeding of Brassica napus L. self-incompatibility restorer lines (1) In spring of 2005 in the test field of Huazhong Agricultural University in Wuhan, and in summer of 2005 in the testing field of Northwest China Testing Station of Huazhong Agricultural University in Hezheng county in Gansu province, Brassica napus L. self-incompatible line S-1300 as female parent was respectively crossed with self-compatible Brassica napus L. varieties Zheyou 18, Huyou 17 and Hui 5900 to obtain seeds of three F1 combinations; in autumn of 2005 the F1 combinations was sowed (3 rows /area, 10 plants /row) in the rapeseed testing field of Huazhong Agricultural University in Wuhan, and the seed-setting circumstances of each combination were investigated in spring of 2006, it was found that the compatibility index of each of the three F1 combinations was more than 10.
This indicated that all the male parents of the F1 combinations can restore the self-incompatibility of Brassica napus L. self-incompatible line S-1300.
(2) In autumn of 2005, the 3 portions of seeds of self-compatible Brassica napus L. were sowed in a randomized block design (2 repeats, 3 rows /area, 10 plants / row) in the rapeseed testing field of Huazhong Agricultural University; the field management is the same as that of the local large field crop production.
(3) Plants of the three kinds of Brassica napus L. were observed to exhibit good field performance, and the compatibility index of the combination between the three kinds of Brassica napus L. and Brassica napus L. self-incompatible line S-1300 was more than 10; This indicated that they can be used as Brassica napus L. self-incompatibility restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900, for further combination and comparative experiments.
4. Breeding of Brassica napus L. self-incompatibility two-line hybrid (1) In spring of 2007, Brassica napus L. self-incompatible line S-1300 and 07-P63-6 YU 34 as female parent were crossed with 3 Brassica napus L.
self-incompatibility restorer lines, respectively (Zheyou 18, Huyou 17 and Hui 5900) to obtain F1 combinations.
(2) In autumn of 2007, the above F1 combinations and one control variety (Zhongyouza No. 2) were sowed in a randomized block design (2 repeats, 3 rows /area, 10 plants / row) in the rapeseed testing field of Huazhong Agricultural University; the field management is the same as that of the local large field crop production.
(3) In May 2008, 10 plants were randomly selected from each area, their agronomic traits such as plant height, branching part and t yield per plan etc were investigated, and the other plants were harvested to investigate the yield per area, and finally the quality traits were analyzed. Through the above steps, the yield per area of the FI combinations according to the invention was increased by 8-12% than that of the control variety Zhongyouza No. 2 (bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, an excellent rapeseed variety popularized widely over China). This indicated that the Brassica napus L. self-incompatibility hybrids have strong heterosis.
5. Propagation of Brassica napus L. self-incompatible lines (1) Brassica napus L. self-incompatible line S-1300 was sowed every year from later August to early September under isolated conditions with spacing in the rows of 2 in and spacing between plants of 1 in in winter rapeseed planting areas along the middle and lower reaches of Yangtze river, China, and Brassica napus L. self-incompatible line with large plant shape and branches as many as possible was bred. Each plant of Brassica napus L.
self-incompatible line S-1300 was encased with a 60-mesh nylon breeze tent at bolting stage, and honeybee pollination was deployed in the tent. The main inflorescences of the self-incompatible line were selfed in bags, and self-compatibility of the plant of the self-incompatible line was identified.
(2) During flowering period, the plants of Brassica napus L.
self-incompatible line S-1300 under isolated condition were sprayed one time every morning with 3% NaCI solution, and sprayed 20-30 times in total.
(3) At mature stage of Brassica napus L. self-incompatible line S-1300, seeds of the self-incompatible line of individual plant were harvested. The compatibility index was calculated by the formula: compatibility index =
seed number / flower number, and the plants whose compatibility index is over 2 were weeded out. Contents of erucic acid and glucosinolates of the seeds of each plant were analyzed, and the seeds with erucic acid content of less than 1% and glucosinolates content of less than 30umol/g were selected as breeder seeds of Brassica napus L. self-incompatible line, and the seeds were kept in refrigerator at -20 C for subsequent use.
(4) The breeder seeds obtained from step (3) were continuously sowed in the next propagation season. In flowering period, plants of the self-incompatible line were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total. Honeybee pollination was deployed, and original seeds of Brassica napus L. self-incompatible line were harvested for use in agricultural production.
6. Propagation method for Brassica napus L. self-incompatibility restorer lines The method is the same as propagation methods of restorer lines including utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, or genic male sterility, or chemical hybridization. On September 25, 2006, Brassica napus L. self-incompatibility restorer line Zheyou 18 was sowed in the seedling bed, and the plants were transplanted in an isolated shelter No. P63-8 on November 5; in 2007 the blossoming flowers were removed in primary flowering stage of Brassica napus L., and the shelter was encased with a 60-mesh nylon breeze tent to isolate the foreign pollen, and honeybee pollination was deployed in the tent; the seeds of Brassica napus L.
self-incompatibility restorer line Zheyou 18 in the isolated shelter were harvested together in mature stage.
7. Seed production method for Brassica napus L. self-incompatibility two-line hybrid (1) In autumn of 2003, Brassica napus L. self-incompatible line S-1300 (female parent) and Brassica napus L. self-incompatibility restorer line Zheyou 18 (male parent) were respectively sowed in the seedling beds; in November 2003, the female parent (S-1300) and male parent (Zheyou 18) were transplanted in a row ratio of 2:1 in the isolated shelter No. P63-2; the field management is the same as that of the local large field crop production.
In 2004 honeybee pollination was deployed in the isolated shelter No. P63-2 in flowering period of Brassica napus L., at the end of final flowering stage, the male parent plants were removed and the female parent plants were kept;
the female parent plants were harvested in mature stage of Brassica napus L.
to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
(2) In autumn of 2007, Brassica napus L. self-incompatible line S-1300 (female parent) and Brassica napus L. self-incompatibility restorer line Hui Zhongshuang No. 10 (male parent) were respectively planted in a row ratio of 2:1 in an isolated area; the field management is the same as that of the local large field crop production. In 2008 at the end of final flowering stage of Brassica napus L., the male parent plants were removed and the female parent plants were kept; the female parent plants were harvested in mature stage of Brassica napus L. to obtain seeds of Brassica napus L.
self-incompatibility two-line hybrid.
(3) The F1 combinations which met the breeding objectives were namely the strong heterosis combinations available for the next experiment. Through the above steps, the strong heterosis combinations according to the invention increased the production by 8-12% than that of the contrast variety Zhongyouza No. 2 (an excellent rapeseed variety bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, approved and popularized widely over China).
4. Propagating of Brassica napus L. self-incompatible lines, which comprises the following steps:
(1) Brassica napus L. self-incompatible line S-1300 was sowed every year from later August to early September under isolated conditions with spacing in the rows of 2 m and spacing between plants of I m in winter rapeseed-planting areas along the middle and lower reaches of Yangtze river, China, and Brassica napus L. self-incompatible line with large plant shape and branches as many as possible was bred. Each plant of Brassica napus L.
self-incompatible line S-1300 was encased with a 60-mesh nylon breeze tent at bolting stage, and honeybee pollination was deployed in the tent. The main inflorescences of the self-incompatible line were selfed in bags, and self-compatibility of the plant of the self-incompatible line was identified.
(2) During flowering period, the plants of Brassica napus L.
self-incompatible line S-1300 under isolated condition were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total.
(3) At mature stage of Brassica napus L. self-incompatible line S-1300, seeds of the self-incompatible line of individual plant were harvested. The compatibility index was calculated by the formula: compatibility index =
seed number / flower number, and the plants whose compatibility index is over 2 were weeded out. Contents of erucic acid and glucosinolates of the seeds of each plant were analyzed, and the seeds with erucic acid content of less than 1% and glucosinolates content of less than 30umol/g were selected as breeder seeds of Brassica napus L. self-incompatible line, and the seeds were kept in refrigerator at -20 C for subsequent use.
(4) The breeder seeds obtained from step (3) were continuously sowed in the next propagation season. In flowering period, plants of the self-incompatible line were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total. Honeybee pollination was deployed, and original seeds of Brassica napus L. self-incompatible line were harvested for use in agricultural production.
5. Propagation of Brassica napus L. self-incompatibility restorer lines:
Propagation methods of restorer lines by utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, or genic male sterility, or chemical hybridization were used to propagate the Brassica napus L.
self-incompatibility restorer lines (see: Breeding and utilization of rapeseed hybrid, page 145-150, editor in chief, FU Tingdong; Hubei Science and Technology Press, 1995) .
6. Seed production method for Brassica napus L. self-incompatibility two-line hybrid:
(1) Brassica napus L. self-incompatible line as female parent was crossed with Brassica napus L. self-incompatibility restorer line as male parent, wherein the female parent and male parent plants were planted under artificial isolated shelter or in natural isolated area in a row ratio of 2:1 or 3:1 or 4:1. The field management is the same as that of the local large field crop production.
(2) In blossoming of the plants of Brassica napus L., honeybees were arranged to assist pollination in artificial isolated shelter or natural isolated area.
(3) At the end of final flowering stage of Brassica napus L., the plants of male parent rows were removed.
(4) At mature stage of Brassica napus L., the female parent row plants were harvested to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
Example 2 (comparative test) The detail steps are as follows:
1. Using compatibility index to evaluate the self-compatibility of Brassica napus L.
At primary flowering stage, the main branches of the investigated Brassica napus L. plants were encased in bags. When there were 3-5 flowers blossoming at a main branch, all the blossoming flowers at the main branch were removed and some little flower buds at the top of inflorescence center were thinned out;
then the main branch and two side branches were encased in a sulfate paper bag and the bag was clipped tightly with paper clips. The paper bag was pulled upward every two days, while patting it to shed the pollen to assistant pollination.
If there were flowers or flower buds revealing out of the bag when the bags were pulled up, the flowers or flower buds should be removed immediately to avoid harvesting impure seeds. When the pollination of almost all flowers in bags was finished, the bags were removed and the buds and un-pollinated flowers were stricken off, a preliminary investigation of self-compatibility was carried out based on the development of rapeseed pods. After harvesting, the seeds were threshed and the compatibility index of each plant was investigated. The compatibility index of a plant was calculated by the formula: compatibility index (Self-compatibility Index, SCI) = seed number /flower number.
2. Breeding of Brassica napus L. self-incompatible line 07-P63-6 YU 34 (1) In spring of 1998, the self-incompatible line S-1300 as female parent was crossed with Hua Shuang No. 3, a semi-winter type Brassica napus L.
line in China, to obtain seeds of the first generation of hybrid (F1), in a rapeseed testing field of Huazhong Agricultural University, Wuhan in Hubei province, China.
(2) In autumn of 1999, F1 seeds were sowed (2 rows, 10 plants /row). In spring of 2000, the F1 plants were selfed in bags in blossoming to evaluate the self-compatibility of F1, and the F1 plants were selfed by artificially bud-pollination. F1 plants were harvested in early May, and the selfed F1 plants hardly went to seed.
(3) In autumn of 2000, the seeds of the selfed F1 plants were sowed to obtain F2 plants (Field number: 01-9-701, 2 rows /area, 10 plants /row, 20 F2 plants in total), in rapeseed testing field of Huazhong Agricultural University, Wuhan; in spring of 2001, 8-10 individual plants of the Brassica napus L. were selected to be selfed and selfed by bud-pollination in blossoming; when the rapeseed pods were mature in early May, the seeds of each selfed F2 plant were harvested separately, and the compatibility index was investigated, the seeds of the plant whose compatibility index is less than 1 were kept.
(4) In 2006, the Brassica napus L. self-incompatible line with stable self-incompatibility and double-low quality was obtained through the breeding method of steps (3)-(6) according to the invention.
In autumn of 2006, the seeds harvested from an individual plant with low compatibility index, low erucic acid content and low glucosinolates content, and more seeds obtained by bud-pollination, were sowed in testing field in Huazhong Agricultural University, Wuhan, and the testing number is YU 34; the plants were transplanted in an isolated shelter No. P63-6 in early November, and the field number is 07-P63-6 YU 34 , the plants were transplanted in 5 rows and has a number of about 50 in total; in spring of 2007, the shelter was encased with nylon breeze tent to isolate the foreign pollen; all the individual plants grew normally, the number of seeds by selfing was small, compatibility index was less than 1, the content of erucic acid <1%, and the content of glucosinolates <30umol/g, they met the double-low standard.
3. Breeding of Brassica napus L. self-incompatibility restorer lines (1) In spring of 2005 in the test field of Huazhong Agricultural University in Wuhan, and in summer of 2005 in the testing field of Northwest China Testing Station of Huazhong Agricultural University in Hezheng county in Gansu province, Brassica napus L. self-incompatible line S-1300 as female parent was respectively crossed with self-compatible Brassica napus L. varieties Zheyou 18, Huyou 17 and Hui 5900 to obtain seeds of three F1 combinations; in autumn of 2005 the F1 combinations was sowed (3 rows /area, 10 plants /row) in the rapeseed testing field of Huazhong Agricultural University in Wuhan, and the seed-setting circumstances of each combination were investigated in spring of 2006, it was found that the compatibility index of each of the three F1 combinations was more than 10.
This indicated that all the male parents of the F1 combinations can restore the self-incompatibility of Brassica napus L. self-incompatible line S-1300.
(2) In autumn of 2005, the 3 portions of seeds of self-compatible Brassica napus L. were sowed in a randomized block design (2 repeats, 3 rows /area, 10 plants / row) in the rapeseed testing field of Huazhong Agricultural University; the field management is the same as that of the local large field crop production.
(3) Plants of the three kinds of Brassica napus L. were observed to exhibit good field performance, and the compatibility index of the combination between the three kinds of Brassica napus L. and Brassica napus L. self-incompatible line S-1300 was more than 10; This indicated that they can be used as Brassica napus L. self-incompatibility restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900, for further combination and comparative experiments.
4. Breeding of Brassica napus L. self-incompatibility two-line hybrid (1) In spring of 2007, Brassica napus L. self-incompatible line S-1300 and 07-P63-6 YU 34 as female parent were crossed with 3 Brassica napus L.
self-incompatibility restorer lines, respectively (Zheyou 18, Huyou 17 and Hui 5900) to obtain F1 combinations.
(2) In autumn of 2007, the above F1 combinations and one control variety (Zhongyouza No. 2) were sowed in a randomized block design (2 repeats, 3 rows /area, 10 plants / row) in the rapeseed testing field of Huazhong Agricultural University; the field management is the same as that of the local large field crop production.
(3) In May 2008, 10 plants were randomly selected from each area, their agronomic traits such as plant height, branching part and t yield per plan etc were investigated, and the other plants were harvested to investigate the yield per area, and finally the quality traits were analyzed. Through the above steps, the yield per area of the FI combinations according to the invention was increased by 8-12% than that of the control variety Zhongyouza No. 2 (bred by Research Institute of Oil Plants, Chinese Academy of Agricultural Sciences, an excellent rapeseed variety popularized widely over China). This indicated that the Brassica napus L. self-incompatibility hybrids have strong heterosis.
5. Propagation of Brassica napus L. self-incompatible lines (1) Brassica napus L. self-incompatible line S-1300 was sowed every year from later August to early September under isolated conditions with spacing in the rows of 2 in and spacing between plants of 1 in in winter rapeseed planting areas along the middle and lower reaches of Yangtze river, China, and Brassica napus L. self-incompatible line with large plant shape and branches as many as possible was bred. Each plant of Brassica napus L.
self-incompatible line S-1300 was encased with a 60-mesh nylon breeze tent at bolting stage, and honeybee pollination was deployed in the tent. The main inflorescences of the self-incompatible line were selfed in bags, and self-compatibility of the plant of the self-incompatible line was identified.
(2) During flowering period, the plants of Brassica napus L.
self-incompatible line S-1300 under isolated condition were sprayed one time every morning with 3% NaCI solution, and sprayed 20-30 times in total.
(3) At mature stage of Brassica napus L. self-incompatible line S-1300, seeds of the self-incompatible line of individual plant were harvested. The compatibility index was calculated by the formula: compatibility index =
seed number / flower number, and the plants whose compatibility index is over 2 were weeded out. Contents of erucic acid and glucosinolates of the seeds of each plant were analyzed, and the seeds with erucic acid content of less than 1% and glucosinolates content of less than 30umol/g were selected as breeder seeds of Brassica napus L. self-incompatible line, and the seeds were kept in refrigerator at -20 C for subsequent use.
(4) The breeder seeds obtained from step (3) were continuously sowed in the next propagation season. In flowering period, plants of the self-incompatible line were sprayed one time every morning with 3% NaCl solution, and sprayed 20-30 times in total. Honeybee pollination was deployed, and original seeds of Brassica napus L. self-incompatible line were harvested for use in agricultural production.
6. Propagation method for Brassica napus L. self-incompatibility restorer lines The method is the same as propagation methods of restorer lines including utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, or genic male sterility, or chemical hybridization. On September 25, 2006, Brassica napus L. self-incompatibility restorer line Zheyou 18 was sowed in the seedling bed, and the plants were transplanted in an isolated shelter No. P63-8 on November 5; in 2007 the blossoming flowers were removed in primary flowering stage of Brassica napus L., and the shelter was encased with a 60-mesh nylon breeze tent to isolate the foreign pollen, and honeybee pollination was deployed in the tent; the seeds of Brassica napus L.
self-incompatibility restorer line Zheyou 18 in the isolated shelter were harvested together in mature stage.
7. Seed production method for Brassica napus L. self-incompatibility two-line hybrid (1) In autumn of 2003, Brassica napus L. self-incompatible line S-1300 (female parent) and Brassica napus L. self-incompatibility restorer line Zheyou 18 (male parent) were respectively sowed in the seedling beds; in November 2003, the female parent (S-1300) and male parent (Zheyou 18) were transplanted in a row ratio of 2:1 in the isolated shelter No. P63-2; the field management is the same as that of the local large field crop production.
In 2004 honeybee pollination was deployed in the isolated shelter No. P63-2 in flowering period of Brassica napus L., at the end of final flowering stage, the male parent plants were removed and the female parent plants were kept;
the female parent plants were harvested in mature stage of Brassica napus L.
to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
(2) In autumn of 2007, Brassica napus L. self-incompatible line S-1300 (female parent) and Brassica napus L. self-incompatibility restorer line Hui Zhongshuang No. 10 (male parent) were respectively planted in a row ratio of 2:1 in an isolated area; the field management is the same as that of the local large field crop production. In 2008 at the end of final flowering stage of Brassica napus L., the male parent plants were removed and the female parent plants were kept; the female parent plants were harvested in mature stage of Brassica napus L. to obtain seeds of Brassica napus L.
self-incompatibility two-line hybrid.
Claims (4)
1. A breeding method for Brassica napus L. self-incompatibility two-line hybrid, comprising the following steps:
(1) breeding of Brassica napus L. self-incompatible line: Brassica napus L. self-incompatible line S-1300 as female parent was crossed with Brassica napus L. self-compatible line Hua Shuang No. 3 to obtain F1 seeds;
the self-compatibility of F1 plants were measured by compatibility index method, and the self-incompatible F1 plants were subjected to selfing by bud-pollination to obtain F2; the self-compatibility of F2 plants were measured, and the self-incompatible F2 plants were selected and subjected to selfing by artificial bud-pollination to obtain F3 seeds; F3 seeds were sowed and the F3 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-stripping to obtain F4 seeds; F4 seeds from F3 family lines in which the compatibility index of individual plant is less than 1 were sowed and the F4 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-pollination to obtain F5 seeds, the F5S from F4 family lines with compatibility index of less than 1 were selected as the desired Brassica napus L. self-incompatible lines; the F3 family lines with unstable self-incompatibility were repeatedly subjected to the above steps until a Brassica napus L. self-incompatible line with stable self-incompatibility was selected; the Brassica napus L. self-incompatible line 07-P63-6 YU 34 was finally obtained;
(2) breeding of Brassica napus L. self-incompatibility restorer lines:
Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatible rapeseed line Zheyou 18, Huyou 17 and Hui 5900 to obtain F1 seeds; F1 seeds were sowed and the self-compatibility of F1 was investigated; said self-compatible rapeseed lines were sowed at the same time, and their yield and yield-related traits were investigated, the breeding materials with good field performance, wherein the compatibility index of F1 combination from that breeding material and said Brassica napus L.
self-incompatible line S-1300 was over 10, were selected as target restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900;
(3) breeding of Brassica napus L. self-incompatibility two-line hybrid:
Brassica napus L. self-incompatible line S-1300 and its improved self-incompatible line 07-P63-6 YU 34 as female parents were crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to breed two-line hybrid; F1 combinations and the contrast varieties were sowed in field at the same time according to randomized block design, and agronomic traits, yield, disease resistance, growth period and quality traits of the plants were investigated to select the combinations with strong heterosis for subsequent use;
(4) propagating of Brassica napus L. self-incompatible lines: the means of early sowing, low density cultivation and high fertilizer were used to cultivate strong seedlings of Brassica napus L. self-incompatible lines, 3%
NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination, the compatibility index of each plant was investigated and the seeds of the plant with compatibility index of less than 1 were harvested as breeder seeds of Brassica napus L.
self-incompatible line, the seeds were kept in refrigerator at -20°C
for subsequent use; a small amount of breeder seeds were sowed in the next sowing season, 3% NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination to propagate the self-incompatible lines, and the original seeds of Brassica napus L. self-incompatible lines were obtained;
(5) propagating of Brassica napus L. self-incompatibility restorer lines:
Propagation methods of restorer lines by utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, genic male sterility, or chemical hybridization were carried out to propagate the Brassica napus L.
self-incompatibility restorer lines;
(6) producing the seeds of Brassica napus L. self-incompatibility two-line hybrid: Brassica napus L. self-incompatible line obtained from step (1) was served as female parent, Brassica napus L. self-incompatibility restorer line obtained from step (2) was served as male parent, wherein the female parent and male parent plants were planted under artificial isolated condition or in natural isolated area in a row ratio of 2:1 or 3:1 or 4:1, in blossoming of Brassica napus L. plants, honeybees were used to supplement pollination, and at the end of final flowering stage of Brassica napus L. the plants of male parent rows were removed, the female parent row plants were harvested at mature stage of Brassica napus L. to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
(1) breeding of Brassica napus L. self-incompatible line: Brassica napus L. self-incompatible line S-1300 as female parent was crossed with Brassica napus L. self-compatible line Hua Shuang No. 3 to obtain F1 seeds;
the self-compatibility of F1 plants were measured by compatibility index method, and the self-incompatible F1 plants were subjected to selfing by bud-pollination to obtain F2; the self-compatibility of F2 plants were measured, and the self-incompatible F2 plants were selected and subjected to selfing by artificial bud-pollination to obtain F3 seeds; F3 seeds were sowed and the F3 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-stripping to obtain F4 seeds; F4 seeds from F3 family lines in which the compatibility index of individual plant is less than 1 were sowed and the F4 family lines were selected, all individual plants of the selected lines were selfed in bags and 3-5 individual plants selected from every line were subjected to selfing by artificial bud-pollination to obtain F5 seeds, the F5S from F4 family lines with compatibility index of less than 1 were selected as the desired Brassica napus L. self-incompatible lines; the F3 family lines with unstable self-incompatibility were repeatedly subjected to the above steps until a Brassica napus L. self-incompatible line with stable self-incompatibility was selected; the Brassica napus L. self-incompatible line 07-P63-6 YU 34 was finally obtained;
(2) breeding of Brassica napus L. self-incompatibility restorer lines:
Brassica napus L. self-incompatible line S-1300 as female parent was crossed with self-compatible rapeseed line Zheyou 18, Huyou 17 and Hui 5900 to obtain F1 seeds; F1 seeds were sowed and the self-compatibility of F1 was investigated; said self-compatible rapeseed lines were sowed at the same time, and their yield and yield-related traits were investigated, the breeding materials with good field performance, wherein the compatibility index of F1 combination from that breeding material and said Brassica napus L.
self-incompatible line S-1300 was over 10, were selected as target restorer lines, i.e., Hui Zheyou 18, Hui Huyou 17 and Hui 5900;
(3) breeding of Brassica napus L. self-incompatibility two-line hybrid:
Brassica napus L. self-incompatible line S-1300 and its improved self-incompatible line 07-P63-6 YU 34 as female parents were crossed with the self-incompatibility restorer lines Hui Zheyou 18, Hui Huyou 17 and Hui 5900 as male parents to breed two-line hybrid; F1 combinations and the contrast varieties were sowed in field at the same time according to randomized block design, and agronomic traits, yield, disease resistance, growth period and quality traits of the plants were investigated to select the combinations with strong heterosis for subsequent use;
(4) propagating of Brassica napus L. self-incompatible lines: the means of early sowing, low density cultivation and high fertilizer were used to cultivate strong seedlings of Brassica napus L. self-incompatible lines, 3%
NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination, the compatibility index of each plant was investigated and the seeds of the plant with compatibility index of less than 1 were harvested as breeder seeds of Brassica napus L.
self-incompatible line, the seeds were kept in refrigerator at -20°C
for subsequent use; a small amount of breeder seeds were sowed in the next sowing season, 3% NaCl solution was sprayed in blossoming of Brassica napus L. under isolated condition with the aid of honeybee pollination to propagate the self-incompatible lines, and the original seeds of Brassica napus L. self-incompatible lines were obtained;
(5) propagating of Brassica napus L. self-incompatibility restorer lines:
Propagation methods of restorer lines by utilizing heterosis such as Brassica napus L. cytoplasmic male sterility, genic male sterility, or chemical hybridization were carried out to propagate the Brassica napus L.
self-incompatibility restorer lines;
(6) producing the seeds of Brassica napus L. self-incompatibility two-line hybrid: Brassica napus L. self-incompatible line obtained from step (1) was served as female parent, Brassica napus L. self-incompatibility restorer line obtained from step (2) was served as male parent, wherein the female parent and male parent plants were planted under artificial isolated condition or in natural isolated area in a row ratio of 2:1 or 3:1 or 4:1, in blossoming of Brassica napus L. plants, honeybees were used to supplement pollination, and at the end of final flowering stage of Brassica napus L. the plants of male parent rows were removed, the female parent row plants were harvested at mature stage of Brassica napus L. to obtain seeds of Brassica napus L. self-incompatibility two-line hybrid.
2. Use of the method according to claim 1 in utilizing the heterosis of Brassica Napus L.
3. The use according to claim 2, comprising the use in breeding Brassica napus L. self-incompatibility two-line hybrid.
4. Use of the method according to claim 1 in breeding Brassica napus L.
self-incompatibility two-line hybrid.
self-incompatibility two-line hybrid.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2008102369602A CN101422133B (en) | 2008-12-23 | 2008-12-23 | Cabbage type rape self-incompatibility two-line hybrid seed-selection method |
| CN200810236960.2 | 2008-12-23 | ||
| PCT/CN2009/001437 WO2010072048A1 (en) | 2008-12-23 | 2009-12-14 | Breeding method for brassica napus l. self-incompatible two-line hybrid |
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| CA2748306A1 true CA2748306A1 (en) | 2010-07-01 |
| CA2748306C CA2748306C (en) | 2017-08-01 |
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| CA2748306A Expired - Fee Related CA2748306C (en) | 2008-12-23 | 2009-12-14 | Breeding method for brassica napus l. self-incompatibility two-line hybrid |
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| Country | Link |
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| CN (1) | CN101422133B (en) |
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| CN101422133B (en) * | 2008-12-23 | 2011-12-21 | 华中农业大学 | Cabbage type rape self-incompatibility two-line hybrid seed-selection method |
| CN103718945A (en) * | 2012-10-11 | 2014-04-16 | 仲衍种业股份有限公司 | Nucleic male sterility two lines rape breeding method |
| CN103155855B (en) * | 2013-04-02 | 2014-12-03 | 镇江瑞繁农艺有限公司 | Method for improving hybrid seed production yield and quality of common head cabbages |
| CN105638447B (en) * | 2016-03-29 | 2017-09-15 | 湖北利众种业科技有限公司 | Improve the selection of Rapeseed GMS system's yield and seed production purity |
| CN114350655B (en) * | 2017-08-24 | 2023-09-26 | 湖南省作物研究所 | Exogenous radish fragment specific marker and preparation method and application thereof |
| CN107466844B (en) * | 2017-09-13 | 2020-04-21 | 北京市农林科学院 | Method for improving radish hybridization seed setting rate |
| CN107896977B (en) * | 2017-12-11 | 2020-07-28 | 青海省农林科学院 | Method for cultivating rape comprehensive hybrid |
| CN108901834A (en) * | 2018-09-07 | 2018-11-30 | 重庆市农业科学院 | A method of wild cabbage self-incompatibility is overcome using chemical method |
| CN111165337A (en) * | 2018-11-09 | 2020-05-19 | 重庆三峡农业科学院 | Cabbage type rape hybrid pollination method |
| CN109588306A (en) * | 2018-12-26 | 2019-04-09 | 华中农业大学 | The method that S haplotype molecular marking supplementary breeding rape is selfed not affine cenospecies |
| CN109566407A (en) * | 2019-01-28 | 2019-04-05 | 华中农业大学 | A kind of method of compound pollination system breeding Brassica napus hybrid kind |
| CN112493117B (en) * | 2020-12-01 | 2021-12-28 | 湖南省作物研究所 | Breeding method of cluster-growing type rape variety with multiple main stems |
| CN114287339B (en) * | 2021-12-20 | 2022-09-06 | 北京市农林科学院 | Cultivation method of green cauliflower rich in beneficial glucosinolates and low in useless glucosinolates |
| CN114158477A (en) * | 2021-12-24 | 2022-03-11 | 苏州市农业科学院 | Breeding method of male sterile line of purple cabbage |
| CN115024215B (en) * | 2021-12-29 | 2023-07-21 | 甘肃农业大学 | Method for producing hybrid seeds between varieties of strong winter/spring cabbage type rape |
| CN115281074B (en) * | 2022-07-01 | 2023-04-18 | 贵州省油菜研究所 | Breeding method of cabbage type rape |
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| CN1089211C (en) * | 1998-12-29 | 2002-08-21 | 华中农业大学 | Developing method of self-incompatible tine and homonuclear maintainer line of cabbage-type rape |
| CN1307866C (en) * | 2004-09-09 | 2007-04-04 | 中国农业科学院油料作物研究所 | Rape cytoplasmic male sterility + self incompatibility hybrid superiority using method |
| CN101250524B (en) * | 2008-04-07 | 2011-05-11 | 华中农业大学 | Molecule marker of brassica napus self-incompatible maintenance line as well as preparation and uses thereof |
| CN101422133B (en) * | 2008-12-23 | 2011-12-21 | 华中农业大学 | Cabbage type rape self-incompatibility two-line hybrid seed-selection method |
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| CN101422133A (en) | 2009-05-06 |
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| CN101422133B (en) | 2011-12-21 |
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