CN101473791B - Method for breeding spring cabbage stalk and maintaining springness of inbred line - Google Patents
Method for breeding spring cabbage stalk and maintaining springness of inbred line Download PDFInfo
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- CN101473791B CN101473791B CN2009100211035A CN200910021103A CN101473791B CN 101473791 B CN101473791 B CN 101473791B CN 2009100211035 A CN2009100211035 A CN 2009100211035A CN 200910021103 A CN200910021103 A CN 200910021103A CN 101473791 B CN101473791 B CN 101473791B
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Abstract
The invention discloses a propagation method for keeping springness of an inbred line of a spring common head cabbage breed. The method comprises the following steps: selecting external-layer green leaves of mature leaf heads of the spring common head cabbage inbred line in spring, inoculating the leaves on an adventitious bud differentiation induction culture medium according to a certain size for culture, and inoculating adventitious buds to an adventitious root differentiation induction culture medium for culture when the adventitious buds grow to 1.5-2.5cm high to obtain complete regenerated seedlings; then transplanting the regenerated seedlings to culture soil for culture, and permanently planting the seedlings in a field (open field) when the seedlings grow to 5-7 true leaves and are adaptively hardened for 7 days, and by which the survival rate of the seedlings reaches over 95%. The method helps improve the breeding effect of the spring common head cabbage, ensure the spring common head cabbage inbred line to complete a heading stage in spring and agronomic character selection of plants of the spring common head cabbage, maintain the characters (springness) of the selected plants unchanged, ensure the inbred line to complete a life cycle, greatly improve the number of the plants with the same genotype of the inbred line, and proliferate the number of the plants over 50 times.
Description
Technical field
The invention belongs to field of plant variety breeding technology, relate to the method that the cultivation that utilizes a kind of biennial plant tissue realizes crop breed breeding in spring, especially a kind of spring head cabbage breeding keeps the propagation method of inbred line spring habit.
Background technology
Head cabbage (Brassica oleracea.L, Var.capitata L.) belong to the Cruciferae rape belong in a kind of important vegetables, it has characteristics such as best in quality, nutritious, edible various, storage tolerance, countries in the world common cultivated.It is reported that Chinese knot cabbage sowing area reached 93.73 ten thousand hm in 2006
2, occupy critical role in vegetable growing and the year-round supply at home.Head cabbage adaptability is strong, can be divided into spring, summer, Qiu Dengduo season, many batches listings in 1 year, to alleviating the market dull and rush season, satisfying the stable market supply of fresh vegetable anniversary and play key effect; Particularly, impel spring head cabbage area under cultivation to enlarge rapidly in recent years along with improving constantly of consumers living and greatly developing of facility cultivation.Head cabbage is typical green body vernalization type biennial crop, have only the plant nutrition body to reach certain volume, could be through certain low temperature induction by vernalization, thereby can only finish a growth period in 1 year, this is just totally unfavorable for precocious first generation of hybrid kind of the spring of suitable spring cultivation and parent's thereof seed selection, and brought the breeding obstacle, make the spring head cabbage of spring cultivation be difficult to breeding in spring.If the head cabbage parental inbred line of spring cultivation often screens in the fall, the spring habit serious degradation that will cause the spring cultivation kind, spring head cabbage kind spring adaptability weaken, poor as occurring balling, lower temperature resistance, weak light resistance easily, bolting phenomenon in advance also easily takes place.Though scientist has worked out the spring habit that can adopt head cabbage axillalry bud cuttage breeding method reservation in spring parental inbred line, but the axillalry bud cuttage seeding cultivation phase is long, through summer, autumn, is subject to summer high temperature, drought impact, make the cuttage seeding survival rate very low, and cost is also very high; Even survive the seedling autumn culture, also can, plant senesecence long because of seedling age, autumn, disease hazard can cause the plant premature dead, lost the inbred line material of institute's seed selection.This just having a strong impact on the suitable spring cultivation kind of seed selection spring head cabbage breeding process.
A kind ofly normally carry out spring head cabbage breeding in spring, can keep the parental autocopulation of institute's seed selection first generation of hybrid kind to tie up to the performance characteristic (spring habit) that has spring, can guarantee the normal survival of selected inbred line again, or enlarge its reproduction coefficient and increase inbred line plant quantity, inbred line is normally finished life cycle, and this just becomes the focus (innovative point) of those skilled in the art's research.
Summary of the invention
The objective of the invention is to, provide a kind of spring head cabbage breeding to keep the propagation method of inbred line spring habit.This method selects for use the green blade of the ripe leaf-head of head cabbage inbred line as explant first, carries out tissue culture, intends obtaining a kind of regrowth with high inductivity, and keeps the propagation method of former inbred line spring habit.
In order to realize above-mentioned task, the present invention takes following technical solution:
A kind of spring head cabbage breeding keeps the propagation method of inbred line spring habit, it is characterized in that, comprises the following steps:
1) explant is selected
Select normal cultivation in spring spring head cabbage inbred line, the outer green blade of the plant leaf-head that selected spring habit performance is good is cut into 1cm
2About small pieces as the explant of cultured in vitro;
2) induction of indefinite bud
Explant is inoculated on the differentiation adventitious buds inducing culture in the blake bottle, and 25 ℃ of temperature, every day, 12h～14h illumination cultivated under intensity of illumination 2000～2500Lx condition, induced indefinite bud through 20～30 days;
The prescription of described differentiation adventitious buds inducing culture is: add 3% sucrose (quality), 0.8% agar (quality), the 6-benzyladenine (6-BA) of 3.0mg/L and the α-Nai Yisuan (NAA) of 0.4mg/L in the MS medium;
3) root induction of indefinite bud
Treat that indefinite bud grows to 1.5～2.5cm when high, downcut on the adventitious bud inducing root medium that it is changed in the blake bottle, 25 ℃ of temperature from the indefinite bud base portion, every day 10h～12h illumination, cultivate under the condition of intensity of illumination 2000～2500Lx,, obtain regrowth through root induction in 15～20 days;
Described adventitious bud inducing root medium is: the α-Nai Yisuan (NAA) that adds 3% sucrose (quality), 0.8% agar (quality) and 0.5～1.0mg/L in the MS medium.
4) exercise of regrowth and transplanting
To there be the cultivation bottleneck of regrowth to open exercise 3～5 days, from blake bottle, take out regrowth, the medium of flush away root, be transplanted to and impel its slow seedling growth in the polypots that nutrition soil is housed, in the fall, when regrowth had 5～7 true leaves, the adaptability hardening was colonizated in field (open country) after 7 days.
The present invention selects the explant of the green blade of spring head cabbage leaf-head as the plant regeneration cultured in vitro first, guaranteed that spring head cabbage inbred line in the selection of heading stage in spring plant economical character with kept the spring habit of inbred line, has overcome additive method (as cotyledon, hypocotyl inducing culture etc.) and can not carry out the defective that spring habit is selected to keep in the field; Guaranteed the survival of spring head cabbage inbred line, not influenced by unfavorable factors such as summer high temperature, high light, arid, can increase substantially the same genotype plant of inbred line number, it is several more than 50 times to expand numerous strain.
Description of drawings
Fig. 1-the 1st, embodiment spring head cabbage inbred line YCF51-99-835168 leaf-head in the spring maturing stage schemes;
Fig. 1-2 is the selected individual plant figure of embodiment spring head cabbage inbred line YCF51-99-835168 breeding for quality in spring;
Fig. 1-the 3rd, embodiment spring head cabbage inbred line YCF51-99-835168 leaf-head leaf explant figure;
Fig. 2-the 1st, embodiment spring head cabbage inbred line BMP01-88-561336 leaf-head blade (explant) adventitious bud inducing figure;
Fig. 2-the 2nd, the root induction figure of embodiment spring head cabbage inbred line BMP01-88-561336 indefinite bud;
Fig. 2-the 3rd, embodiment spring head cabbage inbred line BMP01-88-561336 regrowth is transplanted culturing pot figure.
The present invention is described in further detail below in conjunction with embodiment that accompanying drawing and inventor provide.
Embodiment
According to technical scheme of the present invention, spring head cabbage breeding keeps the propagation method of inbred line spring habit, specifically follows these steps to carry out:
1) explant is selected
Select normal cultivation in spring spring head cabbage inbred line, the outer green blade of the plant leaf-head that selected spring habit performance is good is cut into 1cm
2About small pieces as the explant of cultured in vitro.
2) differentiation adventitious buds inducing culture preparation
Differentiation adventitious buds inducing culture based formulas is: add 3% sucrose (quality), 0.8% agar (quality), the 6-benzyladenine (6-BA) of 3.0mg/L and the α-Nai Yisuan (NAA) of 0.4mg/L in the MS medium.
3) preparation of adventitious bud inducing root medium
Adventitious bud inducing root culture medium prescription is: the α-Nai Yisuan (NAA) that adds 3% sucrose (quality), 0.8% agar (quality) and 0.5～1.0mg/L in the MS medium.
4) induction of indefinite bud
Explant is inoculated on the differentiation adventitious buds inducing culture in the blake bottle, and 25 ℃ of temperature, every day, 12h～14h illumination cultivated under intensity of illumination 2000～2500Lx condition, induced indefinite bud through 20～30 days (d).
5) root induction of indefinite bud
Treat that indefinite bud grows to 1.5～2.5cm when high, downcut on the adventitious bud inducing root medium that it is changed in the blake bottle from the indefinite bud base portion, 25 ℃ of temperature, every day 10h～12h illumination, cultivate under the condition of intensity of illumination 2000～2500Lx, through (d) root induction in 15～20 days, obtain regrowth.
6) exercise of regrowth and transplanting
To have the cultivation bottleneck of regrowth to open 3～5 days (d) of exercise, and take out regrowth from blake bottle, the medium of flush away root is transplanted to and is impelled its slow seedling growth in the polypots that nutrition soil is housed.In the fall, when regrowth had 5～7 true leaves, the adaptability hardening was colonizated in field (open country) in 7 days after (d).
The preparation method of above-mentioned MS medium sees Li Jun bright " Plant Tissue Breeding study course " (second edition, Beijing: China Agricultyre University Press, 2002,32～38) for details.
Below be the embodiment that the inventor provides: need to prove that these embodiment are some preferred embodiments, the invention is not restricted to these embodiment according to experiment showed.
Embodiment 1:
Present embodiment selects for use morning of male parent (inbred line YCF51-99-835168) of the spring head cabbage kind " Qin sweet 50 " (national variety identification) that applicant's spring head cabbage breeding research chamber newly breeds on behalf of experiment material.
1) explant is selected
In generation morning of spring head cabbage inbred line YCF51-99-835168,, 3 true leaves divided seedling once in propagating seeding in cold frame on December 20, when seedling is long when 6～7 true leaves are arranged, in field planting mid-March at open country.The field is by spring head cabbage outdoor cropping normal management, when the plant strain growth leaf-head is ripe (Fig. 1-1), breeding objective, the good plant (Fig. 1-2) of economical character performance are selected to meet in the field, select the explant (Fig. 1-3) of its leaf-head blade as the plant regeneration cultured in vitro.
2) regeneration plant inducing culture
1. indefinite bud induces
Select the outer green blade of the morning of spring head cabbage inbred line YCF51-99-835168 for use, blade is cut into 1cm for ripe leaf-head
2About little side's sheet, with the sterilization of 75% alcohol (40～50 seconds), 0.1% mercuric chloride (8～10 minutes); Be inoculated in the differentiation adventitious buds inducing culture, the prescription of differentiation adventitious buds inducing culture is: add 3% sucrose (quality), 0.8% agar (quality), the 6-benzyladenine (6-BA) of 3.0mg/L and the α-Nai Yisuan (NAA) of 0.4mg/L in the MS medium.
25 ℃ of temperature, every day, 12h～14h illumination cultivated under intensity of illumination 2000～2500Lx condition, induced indefinite bud through 20～30 days (d).According to the suitable field planting time (the Shaanxi field planting time is late August) of field in autumn (open country), but 1～3 generation of subculture prolong incubation time.Generation morning of spring head cabbage inbred line YCF51-99-835168 in, lure bud rate 86.5%, differentiation rate 128%.
Above-mentioned explant number/total explant number * 100 (following identical) that lure bud rate (%)=induce to sprout; Total bud number of differentiation rate (%)=differentiation/total explant number * 100 (following identical).
2. adventive root induces
Treat that indefinite bud grows to 1.5～2.5cm when high, downcut on the adventitious bud inducing root medium that it is changed in the blake bottle from the indefinite bud base portion, adventitious bud inducing root medium is: the α-Nai Yisuan (NAA) that adds 3% sucrose (quality), 0.8% agar (quality) and 0.5～1.0mg/L in the MS medium.25 ℃ of temperature, every day, 10h～12h illumination cultivated under the condition of intensity of illumination 2000～2500Lx, through (d) root induction in 15～20 days, rooting rate 100%.
3. regrowth is transplanted
To have the cultivation bottleneck of regrowth to open 3～5 days (d) of exercise, and take out regrowth from blake bottle, the medium of flush away root is transplanted to and is impelled its slow seedling growth in the polypots that nutrition soil is housed.In late August, when regrowth had 5～7 true leaves, the adaptability hardening was colonizated in field (open country) in 7 days after (d).Rice shoot survival rate 95%, management later on is same as conventional field (open country) management.
Embodiment 2:
Present embodiment selects for use morning of female parent (inbred line BMP01-88-561336) of the spring head cabbage kind " Qin sweet 50 " (national variety identification) that applicant's spring head cabbage breeding research chamber newly breeds on behalf of experiment material.
1) explant is selected
Cultivate for the field morning of maternal inbred line BMP01-88-561336 and the explant system of selection with morning of male parent inbred line YCF51-99-835168 for (referring to embodiment 1).
2) regeneration plant inducing culture
1. indefinite bud induces
The morning of choosing selected spring head cabbage inbred line BMP01-88-561336 is for the outer green blade of the leaf-head explant as cultured in vitro.Take back the laboratory after gathering blade, earlier with flowing water flushing 3 times, again in ultra-clean work with 75% alcohol-pickled 50 seconds (s), 0.1% mercuric chloride sterilization 10 minutes (min), aseptic water washing 3 times, after blade being cut into the square dice that the length of side is 1cm then, blade back down, be inoculated in the differentiation adventitious buds inducing culture, differentiation adventitious buds inducing culture based formulas is to add 3% sucrose (quality), 0.8% agar (quality), the 6-benzyladenine (6-BA) of 3.0mg/L and the α-Nai Yisuan (NAA) of 0.4mg/L in the MS medium.25 ℃ of temperature, every day, 12h～14h illumination cultivated under intensity of illumination 2000～2500Lx condition, induced indefinite bud through 20～30 days (d).According to the suitable field planting time (the Shaanxi field planting time is late August) of field in autumn (open country), but 1～3 generation of subculture prolong incubation time.Generation morning of spring head cabbage inbred line BMP01-88-561336 in, lure bud rate 91.5%, differentiation rate 135% (Fig. 2-1).
2. adventive root induces
Treat that indefinite bud grows to 1.5～2.5cm when high, downcut on the adventitious bud inducing root medium that it is changed in the blake bottle from the indefinite bud base portion, adventitious bud inducing root medium is to add the α-Nai Yisuan (NAA) of 3% sucrose (quality), 0.8% agar (quality) and 0.5～1.0mg/L in the MS medium.25 ℃ of temperature, every day, 10h～12h illumination cultivated under the condition of intensity of illumination 2000～2500Lx, through (d) root induction in 15～20 days, rooting rate 100%.
3. regrowth is transplanted
To have the cultivation bottleneck of regrowth to open 3～5 days (d) of exercise, and take out regrowth from blake bottle, the medium of flush away root is transplanted to and is impelled its slow seedling growth in the polypots that nutrition soil is housed.In late August, when regrowth has 5～7 true leaves (Fig. 2-3), the adaptability hardening was colonizated in field (open country) in 7 days after (d).Rice shoot survival rate 97%, management later on is same as conventional field (open country) management.
Claims (1)
1. the propagation method of a spring head cabbage breeding maintenance inbred line spring habit is characterized in that, comprises the following steps:
1) explant is selected
Select normal cultivation in spring spring head cabbage inbred line, the outer green blade of the plant leaf-head that selected spring habit performance is good is cut into 1cm
2About small pieces as the explant of cultured in vitro;
2) induction of indefinite bud
Explant is inoculated on the differentiation adventitious buds inducing culture in the blake bottle, and 25 ℃ of temperature, every day, 12h～14h illumination cultivated under intensity of illumination 2000～2500Lx condition, induced indefinite bud through 20～30 days;
The prescription of described differentiation adventitious buds inducing culture is: on the basis of MS medium, add sucrose, the agar of mass fraction 0.8%, the 6-benzyladenine (6-BA) of 3.0mg/L and the α-Nai Yisuan (NAA) of 0.4mg/L of mass fraction 3%;
3) root induction of indefinite bud
Treat that indefinite bud grows to 1.5～2.5cm when high, downcut on the adventitious bud inducing root medium that it is changed in the blake bottle, 25 ℃ of temperature from the indefinite bud base portion, every day 10h～12h illumination, cultivate under the condition of intensity of illumination 2000～2500Lx,, obtain regrowth through root induction in 15～20 days;
Described adventitious bud inducing root medium is: the sucrose, the agar of mass fraction 0.8% and the α-Nai Yisuan (NAA) of 0.5～1.0mg/L that add mass fraction 3% in the MS medium;
4) exercise of regrowth and transplanting
To there be the cultivation bottleneck of regrowth to open exercise 3～5 days, from blake bottle, take out regrowth, the medium of flush away root, be transplanted to and impel its slow seedling growth in the polypots that nutrition soil is housed, in the fall, when regrowth had 5～7 true leaves, the adaptability hardening was colonizated in the field open country after 7 days.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101836590B (en) * | 2010-06-10 | 2012-05-23 | 中国农业大学 | Proliferation method of broccoli |
| CN102805035A (en) * | 2012-08-28 | 2012-12-05 | 邢台市蔬菜种子公司 | Common head cabbage tissue culture method |
| CN103125398B (en) * | 2013-03-20 | 2016-02-17 | 江苏省江蔬种苗科技有限公司 | A kind of method for tissue culture improving head cabbage in-vitro regeneration efficiency |
| CN103749127B (en) * | 2014-01-10 | 2015-10-28 | 西北农林科技大学 | The transplanting method of a kind of wild cabbage microspore seedling from test tube to field |
| CN107787836A (en) * | 2016-08-31 | 2018-03-13 | 辽宁东亚农业发展有限公司 | A kind of spring Chinese Cabbage material system breeding technique |
| CN108719062A (en) * | 2018-05-11 | 2018-11-02 | 广州禾立田生物科技有限公司 | A kind of quick breeding method for tissue culture of brussels sprout |
| CN109169278B (en) * | 2018-09-18 | 2021-12-10 | 江苏省农业科学院 | Tissue culture method for improving in-vitro regeneration efficiency of broccoli |
| CN109169279B (en) * | 2018-09-18 | 2022-02-15 | 江苏省农业科学院 | Method for efficiently obtaining regenerated plants by culturing common head cabbage seed pods |
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Application publication date: 20090708 Assignee: Yangling New Huafu Modern Agricultural Co., Ltd. Assignor: Northwest A & F University Contract record no.: 2012610000140 Denomination of invention: Method for breeding spring cabbage stalk and maintaining springness of inbred line Granted publication date: 20110323 License type: Exclusive License Record date: 20120824 |
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