CN112438199B - Breeding method of rape variety suitable for balcony cultivation - Google Patents
Breeding method of rape variety suitable for balcony cultivation Download PDFInfo
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- CN112438199B CN112438199B CN202011506078.2A CN202011506078A CN112438199B CN 112438199 B CN112438199 B CN 112438199B CN 202011506078 A CN202011506078 A CN 202011506078A CN 112438199 B CN112438199 B CN 112438199B
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
Abstract
The invention discloses a breeding method of rape varieties suitable for balcony cultivation, which comprises the steps of hybridizing a cabbage type rape line 167B serving as a female parent with a flower kale, and carrying out multi-generation backcross with hybrid offspring by taking 167B as a male parent until a selfing group with stable and consistent specific characters is obtained; dark culturing the seeds, reserving the young embryo with the hypocotyl shorter than 3.0cm, culturing until the young embryo becomes seedling, selecting the single plant with the height of less than 70cm at the initial flowering stage for multi-generation selfing until obtaining homozygous and stable short-stalk selfing line seeds, and obtaining the rape variety suitable for balcony cultivation. The invention introduces the ornamental character of the collard in the seedling stage into the cabbage type rape by utilizing distant hybridization, thereby realizing the directional improvement of the target character of the cabbage type rape; meanwhile, the young embryo directional screening technology is utilized to realize the rapid and accurate screening of the contracted plant types, so that not only are the manpower and material resources saved, but also the variety selection efficiency is improved.
Description
Technical Field
The invention belongs to the technical field of rape breeding, and particularly relates to a method for breeding a rape variety suitable for balcony cultivation based on distant hybridization, radiation mutagenesis, dark culture directional breeding and the like.
Background
Along with the continuous acceleration of the urbanization process in China, the urban scale is larger and larger, more and more buildings with concrete structures are provided, the green space is reduced, and the urban greening is concerned by people more and more. For a good environment in homes, more and more people move green plants into indoor balconies, rooms and roofs. The cabbage type rape has bright color, longer flowering phase and higher ornamental value. The rape is easy to cultivate and manage, the seedling stage can be used for viewing leaves, the flowering stage can be used for viewing flowers, and the rape is a pot flower with potential. Meanwhile, the rape can successfully overwinter in most areas of China, the rape can be used as an urban greening flower and a green plant in winter, and the rape can be used for flower observation in spring, so that the problems of monotonous urban greening landscape and lack of vitality in winter of China are solved. However, the conventional cabbage type rape material has the characteristics of high plant type, easy lodging, easy formation of high seedlings in indoor planting, branching in flowering period, sparse flower and the like, and is not suitable for indoor or balcony potted plants, so that the strain type is urgently needed to be selected and bred, and the strain has other ornamental characteristics suitable for balcony and urban greening cultivation, such as attractive leaves and flower organs, dense branching and flower growing and the like. In addition, the method of selecting the target character individual plants through field planting in the seedling stage or the mature stage is time-consuming and labor-consuming, and the cost of field planting management is high.
Disclosure of Invention
The technical problem to be solved by the invention is to overcome the defects and shortcomings in the background technology and provide a method for breeding rape varieties suitable for balcony cultivation.
In order to solve the technical problems, the technical scheme provided by the invention is as follows:
a method for breeding rape varieties suitable for balcony cultivation comprises the following steps:
(1) hybridizing the cabbage type rape line 167B serving as a female parent with a collard ornamental variety 'flower collard' to obtain F1Plant growing;
(2) subjecting said F to1Plants are used as female parent, 167B is used as male parent for backcross, more than or equal to 10 branches are screened in each generation, and fertility is realizedBackcrossing normal single plants until obtaining single plants with more than or equal to 10 branches, uniformly grown branches, fan-shaped plant types, leaf-round shapes, wavy leaf edges, basal leaves and leaf stalks shorter than 3.0cm, half stem-holding growth of stem stalks and leaves and normal fertility;
(3) carrying out isolated selfing on the single plants obtained in the step (2) until selfing groups with stable and consistent properties are obtained;
(4) dark culturing the seeds of the selfing population obtained in the step (3), keeping young embryos with hypocotyls shorter than 3.0cm, transplanting the young embryos to substrate soil, continuously culturing the young embryos to be seedlings, and transplanting the seedlings to a test field to obtain a population with compact plant types;
(5) and (2) performing multi-generation selfing on a single plant with the plant height of less than 70cm at the early flowering stage in the plant type-contracted group until a homozygous and stable dwarf selfing line seed is obtained, thus obtaining the rape variety (namely the Fengling No. 1) suitable for balcony cultivation.
Elongation of hypocotyl and internode of the stem is often regulated by the same regulation way, and the plant type contracted progeny population also usually shows that elongation of the hypocotyl is inhibited. Therefore, in the screening process of the progeny population with the contracted plant type, the pre-screening is carried out through the hypocotyl phenotype in the seedling stage under the dark culture condition, so that the workload in the later stage can be reduced, and the cost is reduced.
In the above breeding method, preferably, in the step (4), the seeds of the selfed population obtained in the step (3) pass through a semi-lethal amount before dark culture60Performing mutagenesis treatment on Co gamma ray to obtain M0And carrying out dark culture on the generation seeds. The radiation mutagenesis treatment can shorten the breeding period and improve the breeding efficiency.
More preferably, said semi-lethal amount60The dose of Co gamma ray radiation is 300-1300Gy, more preferably 1100Gy, and the dose rate is 1.2 Gy/min.
Preferably, in the step (4), the temperature of the dark culture is 18-24 ℃, more preferably 22 ℃, and the culture time is 6 d.
Preferably, in the step (5), the rape variety suitable for balcony cultivation has the following characteristics: the leaves are gray green, the leaves are round, the leaf edges are wavy, the leaf stalks of the lower leaves are shorter than 3.0cm, the stem leaves of the mosses are half-embraced to stem and grow, and the petals are round, flat and side-overlapped to grow.
The method overcomes the defects that the conventional cabbage type rape is not suitable for balcony potting and urban greening due to the characteristics of easy formation of high seedlings, easy lodging, sparse flower and branch planting and the like, further expands the multifunctional application field of the cabbage type rape and improves the production benefit of the rape by the steps of distant hybridization, backcrossing, bagging selfing, dark culture and directional breeding and the like. The bred variety Fengling No. 1 has contracted plant types, has the comprehensive advantages of ornamental effect at the seedling stage of the kale and ornamental effect at the flowering stage of the cabbage type rape, and has high ornamental value. The leaves of the variety are gray green, have a leaf round shape, are wavy along the leaves, have the characteristics of extremely short petioles (shorter than 3.0cm) and round and flat petals, are also characterized by being suitable for cultivation in balconies and urban green belts and are used for appreciation.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention combines distant hybridization with a method of directional breeding by dark culture of immature embryos, introduces the ornamental character of the collard in seedling stage into the cabbage type rape by utilizing the distant hybridization, and realizes the directional improvement of the target character of the cabbage type rape; meanwhile, the young embryo directional screening technology is utilized to realize the rapid and accurate screening of the contracted plant types, so that not only are the manpower and material resources saved, but also the variety selection efficiency is improved.
2. The cabbage type rape variety obtained by the breeding method has the hypocotyl length of only 39.1% of 176B in the seedling stage, is not easy to form high-foot seedlings by indoor planting, has the plant height of less than 1.0m in the mature stage, is not easy to fall down, and is suitable for balcony planting and urban greening; meanwhile, the leaves of the variety in the seedling stage are gray green, the leaves are round, the edges of the leaves are wavy, and the petioles of the basal leaves are extremely short (shorter than 3.0cm), so that the variety is suitable for leaf-watching green planting; meanwhile, the flower pot is provided with a plurality of branches which are uniformly distributed, petals are round, flat and side-overlapped, and the flowers are densely planted, so that the flower pot is suitable for viewing areas such as balconies, urban road greenbelts and the like.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to these drawings without creative efforts.
FIG. 1 is a breeding process of 'Fengying No. 1' in the invention;
FIG. 2 is a graph of selection comparison of individuals with suppressed hypocotyl elongation;
fig. 3 is a comparison of 167B and the hypocotyl "syringin 1";
FIG. 4 is the plant morphology observation at the bud stage (a) and the early flowering stage (b) of the Fengling No. 1.
Detailed Description
In order to facilitate understanding of the invention, the invention will be described more fully and in detail with reference to the accompanying drawings and preferred embodiments, but the scope of the invention is not limited to the specific embodiments below.
Unless otherwise defined, all terms of art used hereinafter have the same meaning as commonly understood by one of ordinary skill in the art. The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the scope of the present invention.
Unless otherwise specifically stated, various raw materials, reagents, instruments, equipment and the like used in the present invention are commercially available or can be prepared by existing methods.
Example (b):
the breeding process of the rape variety (Fengling No. 1) suitable for balcony cultivation is shown in figure 1, and comprises the following steps:
(1) implementation material
The cabbage type rape '167B' is a maintainer line of a Borima cytoplasmic male sterile line 167A bred by the Hunan province crop research institute, the whole growth period is about 210 days, the leaf color at the seedling stage is dark green, and 2-3 pairs of cracked leaves have medium growth vigor; the average plant height is 1.63 m, the number of branches is 7-8, the number of siliques per plant is about 253, the number of grains per silique is about 20, and the weight of thousand grains is 3.75 g; lodging resistance and strong disease resistance; erucic acid content 0, thioglycoside content 26.56 micromole/gram (cake), oil content 44.21%;
the 'floral kale' is also called Paeonia suffruticosa, is a variety of brassica oleracea of brassicaceae, and is a 2-year herbaceous plant; is native to Europe, has oval leaves, slightly-notched, dark green outer leaves, multiple and dense branches and even distribution, has red, yellow and white inner leaves which are particularly beautiful, and can be used as a flower bed for viewing in autumn.
(2) Is suitable for breeding BC with ornamental branch habit1Segregating population construction
In early flowering stage of 167B in 2014, 167B unopened flower buds are emasculated, and pollen of kale is taken for pollination to obtain F1Seeds; f in 5 months 20141Planting in artificial climate box to obtain 24 plants F1Selecting 6 strong plants in the initial flowering stage, emasculating unopened flower buds, and pollinating 167B pollen to obtain BC1F1Isolating the population of seeds.
(3) Directional screening of ornamental plant type
Mixing the above BC1Separating the seeds of the colony, sowing the seeds in the test field of Changsha in Hunan in 10 months in 2014, and obtaining 432 single plants in total. The aspects of branching habit, fertility and the like of a group single plant are inspected at the early flowering stage, 9 single plants with more than 10 branches, uniform branches, fan-shaped and leaf-round plant types, wavy leaves, basal leaves and leaf stalks shorter than 3.0cm, half stem clasping of stem leaves of mosses and stems and fertile single plants are obtained; the 9 unopened individual buds are emasculated and then backcrossed with 167B to obtain BC2F1Separating the population of seeds; mixing BC2F1Separating the seeds of the group, sowing the seeds in the Qinghai Xining test field in 2015 for 5 months, repeating the work, and bagging the selected individual plants for selfing; continuously selecting characters such as branching habit, leaf characteristics and the like in selfing offspring, bagging and selfing the selected individual plants, and repeating the breeding process until 10 lines with stable and consistent characters are obtained.
(4) Determination of temperature conditions for directional screening of embryos
Filling 167B seeds on 6 layers of absorbent paper at the bottom of the culture box, and pouring tap water in the culture box; the seeds of the above-harvested 10 lines were sown in 5cm × 5cm square boxes, and 49 seeds were sown in each box. And (3) placing the culture box after sowing in a thermostat for dark culture at the temperature of 18 ℃, 20 ℃, 22 ℃ and 24 ℃, setting 3 times of repetition for each treatment, and observing the growth condition of the plants after culturing for 6 days. The results show that under the 22 ℃ culture condition, seedlings grow more vigorously than at 18 ℃ and 20 ℃ and are more robust than at 24 ℃, so that 22 ℃ is selected for the next analysis.
(5) Directional screening of immature embryos
Adopting the culture method to sow the seeds of the harvested 10 strains in 216 culture boxes in total, carrying out dark culture in a thermostat at 22 ℃, and observing the hypocotyl of the seedling after culturing for 6 days; selecting seedlings with hypocotyl shorter than 3.0cm to obtain 879 seedlings with hypocotyl elongation inhibited, and preliminarily judging the plants to be a population with contracted plant types (FIG. 2); the 879 seedlings were transplanted into 50-hole plug trays filled with matrix soil, and the plugs were transplanted into the experimental field after 20 days of greenhouse culture, resulting in 721 survivors.
(6) Stable plant type contraction character
When the plants grow to about 10 true leaves, the shapes of the plants are further observed, the selection standards are gray green leaves, circular leaf tips, wavy leaf edges, extremely short basal leaf stalks and other ornamental characteristics suitable for the seedling stage, and a total of 45 single plants with the excellent characteristics are obtained. In the flowering period, ornamental character selection is further carried out to obtain 11 single plants which are uniformly grown branches, compact in plant type, fan-shaped, less than 1.0m in plant height and round in petal shape; selfing the obtained 11 single plants, planting selfed seeds in cells, and selecting representative single plants in the cells with consistent group phenotypes for isolated selfing seed reservation; repeating the above work until a stable selfing population with consistent phenotype is obtained, namely named as Fengling No. 1.
(7)60Co gamma ray radiation treatment and suitable radiation dose determination
In order to shorten the breeding period and improve the breeding efficiency, the seeds of 10 strains obtained by directional screening in the step (3) can be subjected to directional screening60CoγPerforming ray radiation treatment, and setting radiation doses as 300Gy, 500Gy, 700Gy, 900Gy, 1100Gy and 1300Gy respectively for contrast tests, wherein the dose rates are all 1.2 Gy/min; the germination rate of the treated seeds is related to60The Co gamma ray irradiation dose is increased and decreased gradually, when the irradiation dose reaches 900Gy, the germination rate is 62.31 percent, when the irradiation dose reaches 1100Gy, the germination rate is 48.42 percent, and when the irradiation dose exceeds 1300Gy, the germination rate is lower than 40 percent, so 1100Gy is selected as the optimal irradiation dose for treatment, and the seeds of 10 strains are subjected to radiation treatment. And (4) directionally screening the seeds obtained after the radiation treatment by adopting the cultivation method in the step (4), wherein the directional screening of the immature embryos is the same as the step (5), and the character stability of plant type contraction is the same as the step (6). The rape variety suitable for balcony cultivation can be screened according to the method, and the breeding period is shorter.
(8) Statistic analysis of the special plant type and quality traits of' Fengling No. 1
The 'Fengling No. 1' and the 167B are planted in a flowerpot with the diameter of 25cm and the height of 30 cm. Statistical analysis is carried out on various characters in the seedling stage and the full-bloom stage, and the length of the hypocotyl of the 'Fengling No. 1' cultured in dark for 6 days is 2.72 +/-0.19 cm, and is obviously shorter than 167B (6.95 +/-0.03), the hypocotyl of the 167B is lodging, and the 'Fengling No. 1' is upright (fig. 3 and table 1). The length of the petiole of the longest leaf at the lower part of the Fengyi No. 1 is 2.63 +/-0.24 cm, and is obviously shorter than 167B (7.07 +/-0.27 cm), so that the Fengyi No. 1 is more compact in the seedling stage (figure 4). In the full-bloom stage, the average internode length of the 'Fengling No. 1' is 2.06 +/-0.43 cm, which is obviously shorter than 5.92 +/-0.76 cm of the '167B', while the effective number of branches of the 'Fengling No. 1' is not reduced, the number of branches is 10.30 +/-0.56, which is obviously more than that of the '167B' (8.08 +/-0.98), so that the plant type of the 'Fengling No. 1' is more compact in the full-bloom stage. The number of the small flowers of the main inflorescence of the Bing Feng 1 (64.1 +/-6.56) and the total number of the small flowers of the lateral branches (166.9 +/-47.2), the number of the small flowers of the main inflorescence of the 167B (69.1 +/-7.48) and the total number of the small flowers of the lateral branches (182.9 +/-67.7) are not significantly different, the length of the main inflorescence of the Bing Feng 1 (22.1 +/-2.45) is significantly shorter than that of the 167B (61.5 +/-4.80), the average length of the lateral branches of the Bing Feng 1 (16.41 +/-2.23 cm) is significantly shorter than that of the 167B (48.88 +/-9.40 cm), so that the whole flower of the Bing 1 plant is more dense, and the density of the small flowers of the main inflorescence of the Bing 1 is 2.90 and is significantly higher than that of the 167B (1.12). In the full-bloom stage, the height of the Fengling No. 1 plant is 89.42 +/-6.76 cm, and is only about half of 167B (163.89 +/-9.85 cm) (shown in Table 1). In general, the elongation of the hypocotyl of the 'Fengling No. 1' is inhibited, and the caucasian seedlings are not easy to form; the plant height is obviously reduced, the plant type is contracted, and the plant is not easy to fall down; the leaves are gray green and round, the leaves are wavy along the edges, the branches and the flowers grow densely, the method is suitable for balcony planting and urban greening, has the comprehensive advantages of collard seedling stage appreciation and cabbage type rape flowering stage appreciation, and has high appreciation value.
The quality of the seed of the 'Bingqing' 1 is analyzed by a near infrared analyzer (Matrix-1, Bruker, Germany, OPUS/QUANT5.5software), and the result shows that the oil content of the seed of the 'Bingqing 1' is 45.23%, the content of the thioglycoside is 21.2 mu mol/g, and the content of the erucic acid is 0.0%, thereby meeting the national standard of 'Shuangxiao'.
Table 1: 167B and Fengling No. 1 plant type related traits statistical analysis
P <0.01, t test.
In summary, the breeding method of the rape variety suitable for balcony cultivation (Fengling No. 1) disclosed by the invention comprises the steps of firstly, introducing excellent characteristics suitable for seedling stage appreciation, such as uniformly-grown branches, fan-shaped plant types, leaf-round shapes, leaf edge waves and the like of collard into cabbage type rape through distant hybridization, and further combining radiation mutagenesis and dark culture directional breeding to realize rapid simultaneous breeding of seedling hypocotyl and plant type characters. The bred Fengling No. 1 seedling is not easy to form a tall-footed seedling, the plant shape is contracted, and meanwhile, the seedling stage leaves are gray green, leaf-round, the leaf edges are wavy, the lower part leaves (commonly called long-petiole) are extremely short (shorter than 3.0cm), stem leaves and stem leaves of the moss are half-embraced to stem and grow, the petals are round, flat and side-overlapped to grow and the like, so that the Fengling No. 1 seedling has high ornamental value and can be used as balcony pot culture and urban greening.
Claims (3)
1. A method for breeding rape varieties suitable for balcony cultivation is characterized by comprising the following steps:
(1) hybridizing the cabbage type rape line 167B serving as a female parent with a collard ornamental variety 'flower collard' to obtain F1Plant growing;
(2) subjecting said F to1The plant is used as a female parent, the 167B is used as a male parent for backcross, and single plants with more than or equal to 10 branches and normal fertility are screened in each generation for backcross until single plants with more than or equal to 10 branches, uniformly grown branches, fan-shaped plant types, circular leaf shapes, wavy leaf edges, basal leaf and leaf stalks shorter than 3.0cm, half stem holding of stem leaves of mosses and stems and normal fertility are obtained;
(3) carrying out isolated selfing on the single plants obtained in the step (2) until selfing groups with stable and consistent properties are obtained;
(4) dark culturing the seeds of the selfing population obtained in the step (3), keeping young embryos with hypocotyls shorter than 3.0cm, transplanting the young embryos to substrate soil, continuously culturing the young embryos to be seedlings, and transplanting the seedlings to a test field to obtain a population with compact plant types;
(5) performing multi-generation selfing on a single plant with the height of less than 70cm in the initial flowering period in the plant type-contracted group until homozygous and stable dwarf inbred line seeds are obtained, and obtaining a rape variety suitable for balcony cultivation;
the rape variety suitable for balcony cultivation has the following characteristics: the leaves are gray green, the leaves are round, the leaf edges are wavy, the leaf stalks of the lower leaves are shorter than 3.0cm, the stem leaves of the mosses are half-embraced to stem and grow, and the petals are round, flat and side-overlapped to grow.
2. The selective breeding method according to claim 1, wherein in the step (4), the temperature of the dark culture is 18-24 ℃ and the culture time is 6 d.
3. The selective breeding method according to claim 2, wherein the temperature of the dark culture in step (4) is 22 ℃.
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