CN107980734A - A kind of selection of high-quality strong cultivated silkworm breed variety - Google Patents
A kind of selection of high-quality strong cultivated silkworm breed variety Download PDFInfo
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- 241000255789 Bombyx mori Species 0.000 title claims abstract description 74
- 238000009395 breeding Methods 0.000 claims abstract description 32
- 230000001488 breeding effect Effects 0.000 claims abstract description 29
- 239000000463 material Substances 0.000 claims abstract description 29
- 239000013077 target material Substances 0.000 claims abstract description 29
- 238000001514 detection method Methods 0.000 claims abstract description 22
- 238000009396 hybridization Methods 0.000 claims abstract description 19
- 238000005516 engineering process Methods 0.000 claims description 14
- 108090000623 proteins and genes Proteins 0.000 claims description 12
- 241000382353 Pupa Species 0.000 claims description 10
- 238000000034 method Methods 0.000 claims description 10
- 238000003757 reverse transcription PCR Methods 0.000 claims description 10
- 230000009897 systematic effect Effects 0.000 claims description 8
- 238000003556 assay Methods 0.000 claims description 7
- 108010004889 Heat-Shock Proteins Proteins 0.000 claims description 6
- 230000005059 dormancy Effects 0.000 claims description 6
- 238000009366 sericulture Methods 0.000 claims description 6
- 230000007850 degeneration Effects 0.000 claims description 5
- 230000008859 change Effects 0.000 claims description 4
- 230000007613 environmental effect Effects 0.000 claims description 4
- 240000000249 Morus alba Species 0.000 description 8
- 235000008708 Morus alba Nutrition 0.000 description 8
- 238000004519 manufacturing process Methods 0.000 description 7
- 230000018109 developmental process Effects 0.000 description 5
- 230000012447 hatching Effects 0.000 description 4
- 230000000694 effects Effects 0.000 description 3
- 235000013601 eggs Nutrition 0.000 description 3
- 230000008569 process Effects 0.000 description 3
- 102000002322 Egg Proteins Human genes 0.000 description 2
- 108010000912 Egg Proteins Proteins 0.000 description 2
- 230000032683 aging Effects 0.000 description 2
- 230000000692 anti-sense effect Effects 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000001418 larval effect Effects 0.000 description 2
- 230000013011 mating Effects 0.000 description 2
- 230000017448 oviposition Effects 0.000 description 2
- 230000008775 paternal effect Effects 0.000 description 2
- 230000029264 phototaxis Effects 0.000 description 2
- 230000000384 rearing effect Effects 0.000 description 2
- 230000037303 wrinkles Effects 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 102000002812 Heat-Shock Proteins Human genes 0.000 description 1
- 239000003181 biological factor Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000006866 deterioration Effects 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000007726 management method Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 210000004681 ovum Anatomy 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 230000035882 stress Effects 0.000 description 1
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Abstract
The invention belongs to breeding technical field, discloses a kind of selection of high-quality strong cultivated silkworm breed variety, and centering system target material " Z83 " and Japanese target material " Z84 " are conventionally raised, and obtains with the excellent individual of detection d cube；It is mutual friendship original seed in being obtained to material " E Soviet Unions " and material " Z83 " hybridization, material " 9902B " and material " Z84 " hybridization is obtained into Japanese mutually friendship original seed；Original seed and Japanese mutually friendship original seed hybridization are mutually handed over into by middle system, the tetra hybrid of acquisition is high-quality strong cultivated silkworm breed variety.The invention can reduce workload, the reliability of work efficiency and selection is improved, shortens the selection and breeding cycle, and can guarantee that objective trait is not lost, can in 34 years rapidly and efficiently cultivate that constitution is strong, detection d cube is excellent and new silkworm variety with the how wet ability of high temperature resistance.
Description
Technical field
The invention belongs to breeding technical field, more particularly to a kind of selection of high-quality strong cultivated silkworm breed variety.
Background technology
The requirement of past silkworm and mulberry breed breeding is good quality and high output, is reformed with the adjustment of agricultural structure and supply side
Deepen constantly, agricultural workforce's structure and production management planning are changed, and are especially subject to serious shadow in agroecological environment
Ringing do not solved with climatic deterioration at all under conditions of, the single breeding objective for emphasizing high yield receives query and challenge,
Originally the silkworm and mulberry kind of selection and breeding cannot fully meet the needs of Sericultural production, and improve the comprehensive resistance of silkworm and mulberry kind, ensure
The high-quality highly resistance of silkworm and mulberry kind, becomes the basic demand to silkworm and mulberry kind.In the past, on breeding method of mulberry silkworm, mainly with hybridization,
Based on the conventional breeding methods such as backcrossing, by whole genomic hybridization, the independent recombinant distributed and restructuring acquisition is favourable, according to table
Type character, the selection of more than 10 a generations are possible to obtain excellent kind.And with molecular biology and molecular genetics skill
The fast development of art, traditional breeding method cannot fully meet the demand of sericultural production development, explore and develop new educate
Kind means, innovation silkworm germ plasm resource have seemed more and more urgent.
The cycle length of the method choice of traditional cultivated silkworm breed variety selection and breeding, poor reliability, heavy workload, especially for resistance product
The selection and breeding of kind, the cycle of selection are longer.By under specific breeding condition, utilizing Semiquatitative RT-PCR assay technology for detection heat shock egg
White expression of the gene BmsHSP27.4 in silkworm body, can effectively ensure that the degeneration-resistant character of silkworm, improve efficiency of selection, shorten breeding
The time limit；Strengthen silk quality selection using living pupa and cocoon-reeling technology at the same time, the efficiency of selection to cultivated silkworm breed variety silk quality can be improved, ensure target
Character is not lost.
The content of the invention
The technical problem of solution：The purpose of the present invention is to solve auxiliary using Semiquatitative RT-PCR assay technology and living pupa and cocoon-reeling
Help the technical problem of the high-quality strong cultivated silkworm breed variety of selection technique selection and breeding, there is provided a kind of selection and breeding side of high-quality strong cultivated silkworm breed variety
Method.
The present invention is achieved in that a kind of selection of high-quality strong cultivated silkworm breed variety includes the following steps：
Step 1: be in robustness kind " EH is new " (from Hunan Province Sericulture Science Insitute introduce) for female parent, silk
The good spring of fine quality is that kind " Su Chun " (being introduced from Inst. of Silkworm, Chinese Academy of Agricultural Sciences) is paternal hybrid in using, and is mesh in acquisition
Mark material " E Soviet Unions "；Authorizing within 2008 " auspicious " (9902) in the new silkworm variety flying crane × auspicious cloud passed through with Hubei Province is
Breeding material, oriented cultivation, Systematic selection obtain Japanese target material " 9902B "；" E Soviet Unions " and " 9902B " is artificially being set
Systematic selection is carried out in the special nurturing an environment put, the selection of economical character is carried out during feeding from generation to generation to each, adopts
With expression of the Semiquatitative RT-PCR assay technology for detection heat shock protein gene BmsHSP27.4 in silkworm body, BmsHSP27.4 bases are selected
Because the individual of a large amount expression is reserved seed for planting subculture, the individual with the how moist shape of high temperature resistance is obtained；
Be in Step 2: target material " Z83 " be from Inst. of Silkworm, Chinese Academy of Agricultural Sciences introduce silk it is of fine quality
Spring with multifibres amount kind 873, separates, high temperature and humidity environment directive breeding is formed through system.Japanese target material " Z84 " is therefrom
Silk spring of fine quality that sericulture research institute of Academy of Agricultural Sciences of state introduces with multifibres amount kind 874, separates, high temperature and humidity ring through system
Border directive breeding forms；Centering system target material " Z83 " and Japanese target material " Z84 " are conventionally raised, right
Each carries out the selection of economical character during feeding from generation to generation, and individual of reserving seed for planting uses the method choice silk of living pupa and cocoon-reeling
Matter, obtains with the excellent individual of detection d cube；
Step 3: it is mutual friendship in being obtained to material " Z83 " hybridization obtained by the material " E Soviet Unions " obtained by step 1 and step 2
Original seed, Japanese mutually friendship original seed is obtained by material " Z84 " hybridization obtained by the material " 9902B " obtained by step 1 and step 2；
Step 4: by it is middle system mutually hand over original seed and it is Japanese mutually hand over original seed hybridization, acquisition tetra hybrid E Soviet Union Z83 ×
9902BZ84, i.e., high-quality degeneration-resistant cultivated silkworm breed variety.
Further, it is two changes, four dormancy kind during middle system's target material uses, the Japanese target material is using Japanese
Two change four dormancy kinds.
Further, the selection of the high-quality strong cultivated silkworm breed variety is under the environmental condition artificially set, using semidefinite
The expression of RT-PCR technology detection BmsHSP27.4 genes is measured, selects the individual of a large amount expression to carry out subculture of reserving seed for planting；
Extension increasing sequence primer is as follows：
Sense primer：5 '-TTGGTTCCAAAACACGAACTGCCAC-3 ',
Anti-sense primer：5’-TTCCTGATTTTGCCAGCGTGCGTTG-3’.
Advantages of the present invention and good effect are：
The time spent in cultivated silkworm breed variety selection and breeding, is few, by the application of RT-PCR technology, efficiency of selection can be made to improve 70%-
80%, the utilization of living pupa and cocoon-reeling technology, can make detection d cube efficiency of selection improve 30%-40%, substantially reduce cultivated silkworm breed variety choosing
The cycle educated.Simultaneously because the shortening in selection and breeding cycle, reduce the manpower during breed breeding and material resources consumption.Can be in 3-4
In year rapidly and efficiently cultivate that constitution is strong, detection d cube is excellent and the new silkworm variety with the how wet ability of high temperature resistance, satisfaction
Sericultural production is to the needs of the cultivated silkworm breed variety of Labor-saving, resistance to extensive feeding, and for silk, enterprise provides excellent raw material.
Brief description of the drawings
Fig. 1 is the selection flow chart of high-quality strong cultivated silkworm breed variety Hubei Province silkworm provided in an embodiment of the present invention.
Embodiment
In order to make the purpose , technical scheme and advantage of the present invention be clearer, with reference to embodiments, to the present invention
It is further elaborated.It should be appreciated that the specific embodiments described herein are merely illustrative of the present invention, it is not used to
Limit the present invention.
The selection of Silkworm varieties Hubei Province silkworm No. 6 includes the following steps：
S101, be in robustness kind " EH is new " (from Hunan Province Sericulture Science Insitute introduce) for female parent, detection d cube
The excellent spring is that kind " Su Chun " (being introduced from Inst. of Silkworm, Chinese Academy of Agricultural Sciences) is paternal hybrid in using, and is target in acquisition
Material " E Soviet Unions "；" auspicious " (9902) in the new silkworm variety flying crane × auspicious cloud passed through are authorized for 2008 with Hubei Province to educate
Kind material, oriented cultivation, Systematic selection obtain Japanese target material " 9902B "；" E Soviet Unions " and " 9902B " is artificially being set
Special nurturing an environment in carry out Systematic selection, to each from generation to generation during feeding carry out economical character selection, use
Expression of the Semiquatitative RT-PCR assay technology for detection heat shock protein gene BmsHSP27.4 in silkworm body, selects BmsHSP27.4 genes
The individual of a large amount expression is reserved seed for planting subculture, obtains the individual with the how moist shape of high temperature resistance.
The special nurturing an environment and selection that the different generations is set are as shown in table 1 below：
The special nurturing an environment and selection that table 1 " E Soviet Unions " and " 9902B " different generation are set
S102, middle system's target material " Z83 " are from the silk of Inst. of Silkworm, Chinese Academy of Agricultural Sciences's introduction spring of fine quality
With multifibres amount kind 873, separated through system, high temperature and humidity environment directive breeding forms.Japanese target material " Z84 " is from China
Silk spring of fine quality that sericulture research institute of Academy of Agricultural Sciences introduces with multifibres amount kind 874, separates, high temperature and humidity environment through system
Directive breeding forms；Centering system target material " Z83 " and Japanese target material " Z84 " are conventionally raised, to every
One generation carries out the selection of economical character during feeding, and individual of reserving seed for planting uses the method choice detection d cube of living pupa and cocoon-reeling,
Obtain with the excellent individual of detection d cube.
S103, material " Z83 " hybridization to the material " E Soviet Unions " obtained by S101 and obtained by S102 obtain in be it is mutual hand over original seed,
Material " Z84 " hybridization obtained by material " 9902B " obtained by S101 and S102 is obtained into Japanese mutually friendship original seed.
S104, by it is middle system mutually hand over original seed and it is Japanese mutually hand over original seed hybridization, acquisition tetra hybrid E Soviet Union Z83 ×
9902BZ84, is named as Hubei Province silkworm No. 6, i.e., high-quality degeneration-resistant cultivated silkworm breed variety.
The present invention selects the objective trait of resistance parent by RT-PCR technology, utilizes the silk of living pupa and cocoon-reeling Selection parent
Matter character, can go out high-quality degeneration-resistant new silkworm variety by high effect culture in 3-4.
The present invention is target material in being bred as：
It is mainly characterized by：In be two changes, four dormancy kind.Young silkworm has close property, phototaxis.Each age development is neat, and moultinism is fast,
Molting homogeneous；Big to nibble that Sang Sudu is fast, and build is sturdy, body colour is bluish white, plain spot；Aging to gush together, cocooning is fast, seeks upper strata cocoon.Cocoon shape
Short ellipse, cocoon color is pure white, and contracting wrinkle is medium.Moth appearance is neat, and moth white, spreads the wings, mating production of hybrid seeds performance is excellent.Oviposition is slightly slow, and production is attached good, no
Embryonated egg is few, and ovum amount is more, the single moth egg 566 or so of spring breeding.Spring hastened the hatching of silkworms by 10 days, 24 days or so larval phase, pupa
16 days phases, full period 50 days.With Japanese target material to handing over, preferably postpone outbound in 1 day and hasten the hatching of silkworms.
The Japanese target material that the present invention is bred as：
It is mainly characterized by：Japanese two change four dormancy kinds.Young silkworm has close property, phototaxis.Each instar larvae development is neat, moultinism
It hurry up, molting homogeneous；Mulberry speed is nibbled greatly, build is sturdy, and body colour is bluish white, common spot；Aging to gush together, cocooning is fast, seeks upper strata
Cocoon.Cocoon shape length is ellipse shallower with a tight waist, and cocoon color is pure white, and contracting wrinkle is medium.Moth appearance is neat, and moth white, spreads the wings.Easily mating, oviposition is fast, and production is attached
It is good, the single moth egg 530 or so of spring breeding.Spring hastened the hatching of silkworms by 10 days, and when larval phase 24 days 12 is small, pupa time 16 days 12 is small
When, full period 51 days.With middle system's target material to handing over, preferably 1d outbounds ahead of time are hastened the hatching of silkworms.
Selection and technical finesse below in conjunction with the high-quality strong cultivated silkworm breed variety of embodiment detailed description of the present invention
Process.
1st, using the explanation of material
Existing result of study shows that existing heat shock protein rises important in the growth and development process of silkworm in Silkworm, Bombyx mori
Effect, can and biological factor induced expression abiotic by some.Silkworm BmsHSP27.4 genes are not expressed or expressed at normal temperatures
Horizontal extremely low, the expression of the gene rises rapidly under high temperature impact, shows that the gene belongs to thermo-responsive gene, and the gene
Height expression is that silkworm body adapts to the stress reaction that high temperature produces, to protect body from high-temperature damage.(Jin Xin etc., high temperature (34 DEG C)
Under the conditions of silkworm heat shock protein gene BmsHSP27.4 expression change, sericulture science, 2014,40 (4):0620-0626).This
Invention is under the environmental condition artificially set, using the table of BmsHSP27.4 genes in Semiquatitative RT-PCR assay technology for detection Silkworm, Bombyx mori
Reach, select the individual of a large amount expression to carry out subculture of reserving seed for planting.Based at present, also aided in without such technology applied to silkworm breeding
Selection, accelerates the relevant report of silkworm breeding process.
Extension increasing sequence primer is as follows：
Sense primer：5 '-TTGGTTCCAAAACACGAACTGCCAC-3 ',
Anti-sense primer：5’-TTCCTGATTTTGCCAGCGTGCGTTG-3’
2nd, specific implementation steps：
(1) suitable parent is selected according to " multifibres amount × robustness " character principle of complementarity, prepares " in × in ", " day × day "
The combination of hybridization original seed, to be kind " EH is new " in robustness for female parent, the detection d cube excellent spring use in be that kind " Su Chun " is father
This hybridization, based material " E Soviet Unions " in acquisition, is authorized in the new silkworm variety flying crane × auspicious cloud passed through for 2008 with Hubei Province
" auspicious " (9902) are breeding material, and oriented cultivation, Systematic selection obtain Japanese target material " 9902B "；From " E Soviet Unions " and
The F1 generation of " 9902B " starts, and carries out the high temperature and humidity of continuous multi-generation, closed, and the environmental condition of the rearing with less number of feeds a day is cultivated, and F1-F5 generations are complete
Age temperature is 29 DEG C -32 DEG C, 1-4 ages for prevent entirely it is dry educate, 5 ages for half it is anti-it is dry educate, full age daily 2 does not select back to mulberry, mulberry leaf.F5
The generation condition of later then annual higher 28 DEG C -29 DEG C of the temperature of phase in spring room temperature is cultivated, and so that detection d cube selects, the summer and autumn phase is with 29
DEG C of -30 DEG C high temperature are cultivated, to strengthen the ability of high temperature resistance.In F1-F3 generations, carry out mixing ant amount and educate, emphatically individual choice, and F4 generations adopt
Mated, attached most importance to robustness, area elects main combination individual choice as with moth area with one batch rearing.In the special cultivation ring artificially set
Systematic selection is carried out in border, the selection of economical character is carried out during feeding from generation to generation to each, using Semiquatitative RT-PCR assay
Expression of the technology for detection heat shock protein gene BmsHSP27.4 in silkworm body, selects of BmsHSP27.4 genes a large amount expression
Body is reserved seed for planting subculture, obtains the individual with the how moist shape of high temperature resistance.
(2) centering system target material " Z83 " and Japanese target material " Z84 " are conventionally raised, to each
A generation carries out the selection of economical character during feeding, and individual of reserving seed for planting strengthens the choosing of detection d cube using the method for living pupa and cocoon-reeling
Select, obtain with the excellent individual of detection d cube.
(3) it is that mutual friendship is former in being obtained to material " Z83 " hybridization obtained by the material " E Soviet Unions " obtained by step (1) and step (2)
Kind, Japanese mutually friendship original seed is obtained by material " Z84 " hybridization obtained by the material " 9902B " obtained by (1) and step (2).
(4) original seed and Japanese mutually friendship original seed hybridization, tetra hybrid E Soviet Unions Z83 × 9902B of acquisition are mutually handed over into middle system
Z84, is named as Hubei Province silkworm No. 6, i.e., high-quality strong cultivated silkworm breed variety.
The foregoing is merely illustrative of the preferred embodiments of the present invention, is not intended to limit the invention, all essences in the present invention
All any modification, equivalent and improvement made within refreshing and principle etc., should all be included in the protection scope of the present invention.
Sequence table
<110>Crop Institute, Hunan Academy of Agricultural Sciences
<120>A kind of selection of high-quality strong cultivated silkworm breed variety
<130> 2017
<160> 2
<170> SIPOSequenceListing 1.0
<210> 1
<211> 25
<212> DNA
<213>Artificial sequence (Bombyx mori)
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ttggttccaa aacacgaact gccac 25
<210> 2
<211> 25
<212> DNA
<213>Artificial sequence (Bombyx mori)
<400> 2
ttcctgattt tgccagcgtg cgttg 25
Claims (3)
1. a kind of selection of high-quality strong cultivated silkworm breed variety, it is characterised in that the selection of the cultivated silkworm breed variety includes as follows
Step：
It is that kind " Su Chun " is miscellaneous for male parent in the excellent spring use of detection d cube Step 1: to be that kind " EH is new " is female parent in robustness
Hand over, be target material " E Soviet Unions " in acquisition；With " auspicious " 9902 in new silkworm variety flying crane × auspicious cloud for breeding material, warp
Directive breeding, Systematic selection obtain Japanese target material " 9902B "；By " E Soviet Unions " and " 9902B " in the special cultivation artificially set
Systematic selection is carried out in environment, the selection of economical character is carried out during feeding from generation to generation to each, using sxemiquantitative RT-
Round pcr detects expression of the heat shock protein gene BmsHSP27.4 in silkworm body, selects the expression of BmsHSP27.4 genes a large amount
Individual reserve seed for planting subculture, obtain the individual with the how moist shape of high temperature resistance；
It is that target material " Z83 " was used from the silk of Inst. of Silkworm, Chinese Academy of Agricultural Sciences's introduction spring of fine quality in Step 2:
Multifibres amount kind 873, separates, high temperature and humidity environment directive breeding is formed through system.Japanese target material " Z84 " is from Chinese agriculture
Silk spring of fine quality that sericulture research institute of the industry academy of sciences introduces with multifibres amount kind 874, separates, high temperature and humidity environment is determined through system
Formed to selection and breeding；Centering system target material " Z83 " and Japanese target material " Z84 " are conventionally raised, to each
A generation carries out the selection of economical character during feeding, and individual of reserving seed for planting uses the method choice detection d cube of living pupa and cocoon-reeling, obtains
There must be the excellent individual of detection d cube；
Step 3: it is that mutual friendship is former in being obtained to material " Z83 " hybridization obtained by the material " E Soviet Unions " obtained by step 1 and step 2
Kind, Japanese mutually friendship original seed is obtained by material " Z84 " hybridization obtained by the material " 9902B " obtained by step 1 and step 2；
Step 4: original seed and Japanese mutually friendship original seed hybridization, tetra hybrid E Soviet Unions Z83 × 9902B of acquisition are mutually handed over into middle system
Z84, i.e., high-quality degeneration-resistant cultivated silkworm breed variety.
2. such as the selection of the high-quality strong cultivated silkworm breed variety in claim 1, it is characterised in that middle system's target material is adopted
It is two changes, four dormancy kind in, the Japanese target material is using Japanese two four dormancy kinds of change.
3. such as the selection of the high-quality strong cultivated silkworm breed variety in claim 1, it is characterised in that the high-quality strong silkworm product
The selection of kind is under the environmental condition artificially set, using the table of Semiquatitative RT-PCR assay technology for detection BmsHSP27.4 genes
Reach, select the individual of a large amount expression to carry out subculture of reserving seed for planting.
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| Publication number | Priority date | Publication date | Assignee | Title |
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