CN109006464B

CN109006464B - Simplified production method of rape male sterility hybrid F1 seed

Info

Publication number: CN109006464B
Application number: CN201810928989.0A
Authority: CN
Inventors: 杨光圣; 董发明; 辛强; 洪登峰
Original assignee: WUHAN LIANNONG SEED TECHNOLOGY CO LTD; Huazhong Agricultural University
Current assignee: WUHAN LIANNONG SEED TECHNOLOGY CO LTD
Priority date: 2018-08-15
Filing date: 2018-08-15
Publication date: 2021-12-14
Anticipated expiration: 2038-08-15
Also published as: CN109006464A

Abstract

The invention provides a simplified production method of rape male sterile hybrid F1 seeds, which comprises the steps of selecting a cytoplasmic male sterile line (or a nuclear male sterile line) and a restorer line with self-incompatibility characteristic to produce hybrid F1 seeds, wherein the specific method comprises the steps of mixing the cytoplasmic male sterile line (or the nuclear male sterile line) and the restorer line seeds with self-incompatibility characteristic according to a specific proportion under the isolation condition, sowing the uniformly mixed seeds in a field of a rape seed production area, and mixing the sterile line and the restorer line when the seeds are mature to produce hybrid F1 seeds. The invention has simple breeding program, saves labor cost, money and labor.

Description

Simplified production method of rape male sterility hybrid F1 seed

Technical Field

The invention belongs to the technical field of rape hybrid F1 seed production, and particularly relates to a hybrid seed production method for producing hybrid F1 seeds by using a cytoplasmic male sterile line or a nuclear male sterile line and a restorer line with self-incompatibility characteristics.

Background

The rape hybrid vigor is obvious, the yield can be increased by 20-30% generally, the good combination yield increase amplitude can reach 50%, the popularization and application of the hybrid rape are rapidly developed, and the area of the hybrid rape in China only accounts for more than 70% of the total area of the hybrid rape. Heterosis utilization is an important way to increase crop yield. Rape heterosis utilization approaches are many, and cytoplasmic male sterility, nuclear male sterility, chemical male killing and the like are mainly adopted.

The production method of hybrid F1 seed includes planting sterile line and restoring line in certain row ratio in field, and harvesting hybrid F1 seed when the seed is mature. The method has the disadvantages that the sterile line and the restorer line need to be planted separately according to a certain row ratio during planting, the restorer line needs to be harvested separately during harvesting, or the restorer line is cut off after the restorer line withers flowers, and F1 hybrid seeds are harvested on the sterile line when the seeds are mature. Meanwhile, strict supervision is required in the seed harvesting process to avoid mixing the male parent restorer line seeds into the hybrid F1 seeds knotted on the female parent sterile line. Obviously, the existing method wastes time and labor and has higher labor cost.

Disclosure of Invention

The invention aims to overcome the defects of the prior art and provide a simplified method for producing rape hybrid F1 seeds by using a cytoplasmic male sterile line or a nuclear male sterile line.

In order to realize the purpose of the invention, the inventor finally obtains the following simple and efficient technical scheme by combining the research experience of rape male sterility hybridization for many years:

a simplified method for preparing the hybrid F1 seed from the male sterile line of rape (or nuclear male sterile line) and the restoring line with the characteristic of self-incompatibility includes such steps as mixing the cytoplasmic male sterile line (or nuclear male sterile line) with the restoring line with the characteristic of self-incompatibility, sowing the uniformly mixed seeds in the field of rape seed preparing area, and mixing sterile line with restoring line when it is mature to obtain the hybrid F1 seed.

Further preferably, the cytoplasmic male sterile line (or the nuclear male sterile line) and the restorer line seed with the self-incompatibility characteristic in the above method are 80-90% in a suitable ratio: 10 to 20 percent.

Further preferably, the method for producing the male sterile hybrid rape F1 seed is simplified as described above, wherein the isolation conditions include natural isolation and artificial isolation.

Further preferably, the simplified production method of the rape male sterile hybrid F1 seed is as described above, wherein the artificial broadcast sowing is carried out by 6000-30000 plants per mu, the mechanical or artificial drill sowing is carried out by row spacing of 10-25 cm, and the plant spacing is 5-15 cm.

Still further preferably, the simplified method for producing male sterile hybrid F1 seeds for oilseed rape as described above, wherein the artificial broadcast is planted per mu of 20000-25000 plants, the row spacing of the mechanical or artificial drill is 16-22cm, and the plant spacing is 8-12 cm.

Compared with the prior art, the hybrid seed production method has the advantages that:

(1) when sowing, the traditional method is to plant the sterile line and the restorer line separately according to a certain row ratio, which is labor-consuming and time-consuming; the invention has the advantages of mixed planting of the sterile line and the restorer line, simple breeding procedure, labor cost saving, money saving and labor saving.

(2) During harvesting, the traditional method is to separately harvest the sterile line and the restorer line, or to cut the restorer line in advance; the invention directly mixes and collects the sterile line and the restorer line, has simple operation and further saves the labor cost.

(3) The restorer line has the characteristic of self-incompatibility, so that the restorer line cannot fruit normally, the flowering period is prolonged, pollination is facilitated, and the yield of F1 seeds is increased.

(4) Because the restorer line has the characteristic of self-incompatibility, the restorer line cannot fruit normally, and the risk of mixing the male parent restorer line seed into the F1 seed is reduced fundamentally.

Detailed Description

The present invention is further illustrated by the following specific examples. In the following description, numerous specific details are set forth in order to provide a thorough understanding of the present invention. The present invention may be embodied in many different forms without departing from the spirit or essential characteristics thereof, and it should be understood that various changes in form and details can be made therein by those skilled in the art without departing from the spirit and scope of the invention as defined in the appended claims. The following experimental methods are conventional methods and techniques in the art unless otherwise specified. In addition, in the examples, the cytoplasmic male sterile line 616A is a new variety cultivated by the present inventors, and the right of new variety of plant was obtained in 2018, 1 month and 2 days, and the right of variety is CNA 20141083.8.

Example 1: breeding of balima cytoplasmic male sterility restoring line 7-5SR with self-incompatibility characteristic

(1) Crossing with a self-incompatible single plant (the genotype is BnS-1300BnS-1300) separated from an F2 progeny of Hua YOUZAO 95 by using Hua YOUZAO No. 12 restorer line 7-5 as a female parent (the genotype is bnS-1300bnS-1300) to obtain F1.

(2) Backcrossing 7-5 using F1 as the female parent gave BC 1.

(3) And (3) carrying out DNA molecular detection on the BC1 group single plants by using a DNA molecular marker linked with the self-incompatibility gene BnS-1300, and screening the single plants with heterozygous genotypes (BnS-1300bnS-1300) at the self-incompatibility gene locus.

(4) Backcrossing the heterozygous single plant serving as a male parent with 7-5 to obtain BC 2.

(5) And (3) carrying out DNA molecular detection on the BC2 group single plants by using a DNA molecular marker linked with the self-incompatibility gene BnS-1300, and screening the single plants with heterozygous genotypes (BnS-1300bnS-1300) at the self-incompatibility gene locus.

(6) Backcrossing the heterozygous single plant serving as a male parent with 7-5 to obtain BC 3.

(7) And (3) carrying out DNA molecular detection on the BC3 group single plants by using a DNA molecular marker linked with the self-incompatibility gene BnS-1300, and screening the single plants with heterozygous genotypes (BnS-1300bnS-1300) at the self-incompatibility gene locus.

(8) The hybrid single plants are selfed to obtain BC3F 2.

(9) DNA molecular detection is carried out on the individual plants of the BC3F2 population by using a DNA molecular marker linked with the self-incompatibility gene BnS-1300, and the individual plants with homozygous genotypes (BnS-1300BnS-1300) at the self-incompatibility gene locus are screened out.

(10) The individuals having the homozygous genotypes (BnS-1300BnS-1300) at the self-incompatible gene loci were self-pollinated (artificial pollination) to obtain BC3F 3.

(11) And (3) carrying out agronomic trait and self-incompatibility identification on the BC3F3 family, screening out a family which is self-incompatible and has a phenotype closest to 7-5, and carrying out mixed propagation (artificial auxiliary pollination) mixed harvest to obtain 7-5 SR.

Example 2: cytoplasmic male sterility hybrid seed production for rape

The method comprises the following steps of producing hybrid F1 seeds by using a cytoplasmic male sterile line 616A and a restorer line 7-5SR with self-incompatibility characteristics, wherein the hybrid seed production method comprises the following specific steps:

(1) mixing the cytoplasmic male sterile line 616A and the restorer line 7-5SR seeds with self-incompatibility characteristic according to the proportion of 80%: mixing at a ratio of 20%.

(2) And under the condition of isolating an artificial net room, planting the uniformly mixed seeds according to the row spacing of 20cm and the plant spacing of 10 cm.

(3) When the seeds are mature, the sterile line and the restorer line are harvested in a mixed mode, and statistics shows that the purity of the hybrid is 92.6%.

Example 3: cytoplasmic male sterility hybrid seed production for rape

(1) Mixing 80% of nuclear male sterile line 616A and restorer line 7-5SR seeds with self-incompatibility characteristic: mixing at a ratio of 20%.

(2) Under the condition of natural isolation, the seeds mixed uniformly are planted according to the density of 25000 plants per mu.

(3) When the seeds are mature, the sterile line and the restorer line are harvested in a mixed mode, and statistics shows that the purity of the hybrid is 90.6%.

Example 4: rape nuclear male sterility hybrid seed production

(1) The nuclear male sterile line RG430A and the restorer line 7-5SR seed with self-incompatibility characteristic are mixed according to the proportion of 90%: mixing at a ratio of 10%.

(3) When the seeds are mature, the sterile line and the restorer line are harvested in a mixed mode, and statistics shows that the purity of the hybrid is 97.5%.

Example 5: rape nuclear male sterility hybrid seed production

(2) Under the condition of natural isolation, the seeds which are uniformly mixed are planted according to the density of 20000 plants/mu. .

(3) When the seeds are mature, the sterile line and the restorer line are harvested in a mixed mode, and statistics shows that the purity of the hybrid is 91.8%.

Claims

1. A simplified method for producing male sterile hybrid F1 seed of rape, which comprises selecting cytoplasmic male sterileThe sexual sterile line or the nuclear male sterile line is used as the female parent and hasBnS-1300The restoring line with gene self-incompatibility characteristic is used as male parent, under the condition of isolation the seeds of female parent and male parent are mixed, the uniformly-mixed seeds are sowed in the field of rape seed-producing area, when the seeds are ripe, the sterile line and restoring line are mixed, so that the rape male sterile hybrid F1 seed can be obtained.

2. The simplified method for producing male sterile hybrid rape F1 seed according to claim 1, wherein the ratio of female parent seed to male parent seed is 80-90%: uniformly mixing 10-20% of the mixture and then sowing.

3. The method for producing simplified male sterile hybrid rape F1 seed according to claim 1, wherein the isolation conditions include natural isolation and artificial isolation.

4. The simplified method for producing male sterile hybrid F1 seed of Brassica napus as claimed in claim 1, wherein 6000-30000 plants per mu are planted by manual sowing, the row spacing of mechanical or manual drilling is 10-25 cm, and the row spacing is 5-15 cm.

5. The simplified method for producing male sterile hybrid F1 seed of Brassica napus as claimed in claim 4, wherein the row spacing of mechanical or artificial sowing is 16-22cm and the row spacing is 8-12cm, wherein 20000-25000 plants per mu are planted by artificial sowing.