CN103749401A - Practical silkworm variety cultivation method capable of controlling male ratio and application - Google Patents
Practical silkworm variety cultivation method capable of controlling male ratio and application Download PDFInfo
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Abstract
The invention provides a practical silkworm variety cultivation method capable of controlling the male ratio. In the method, a sex-limited ant color system under the control of an sch mutant gene carried on Z chromosome of silkworm and a +sch gene translocated on a W chromosome is taken as a donor, and a Z<sch> gene and a W<+sch> gene of the donor can transferred onto a receptor through hybridization in which an ordinary ant color practical silkworm variety is taken as the receptor. In the method, the genetic regularity of key genes (namely, the Z<sch> gene and the W<+sch> gene) for controlling the male ratio are organically combined with a classical silkworm breeding method, thereby forming a practical silkworm variety method capable of controlling the male ratio in rapid breeding is constructed.
Description
Technical field
The invention belongs to Perspective of Animal Breeding Methods, particularly silkworm breed of variety.
Technical background
Silkworm is a kind of important economic insects, is human use's historical origin of existing more than 5000 year.Silkworm is to multiply by sexual reproduction, and the individual ratio of offspring's male and female is impartial, forms a kind of balance of nature, is convenient to derive without interruption offspring.Silkworm is female and malely have respectively different features, need to be to female or male control in order to reach certain target in production.
The sex chromosome of silkworm is ZW type, and W chromosome has female decisive action.Scientists utilizes this feature by chromosome engineering, the system such as successively obtain that speckle is sex-limited, ovum look sex-limited and ant look sex-limited, and these all can distinguish male and female according to morphological markers, and expectation realizes the object of regulation and control female-male proportion thus.Speckle is sex-limited need to be arrived the grown silkworm phase and could differentiate male and female, at this moment aspect raising, has dropped into a large amount of manpower and materials, and uneconomic shortcoming is outstanding.Ovum look sex-limited differentiates male and female in the ovum phase, the ant look sex-limited newly-hatched silkworm body colour according to just hatching is differentiated male and female, though this two type systematic has been avoided the excessively late puzzlement bringing of sex-limited resolution phase of speckle to a great extent, but it is very little that male and female are differentiated the individual body profile of phase, from the colony of male and female equal distribution, the difficulty of safe separating male and female individuality is very large, operability is very poor, and manual operations can not adapt to the upper a large amount of demand of using of producing at all.
Silkworm is that sperm and ovum are combined into dliploid and develop into individuality.Male moth of silkworm can produce 1,400,000-2,000,000 enpyrene spermatozoons, and a female moth generally only produces 300-600 grain ovum; Male moth can with a plurality of female moth mating, can make a plurality of female moth normal fertilization.Female proportion shared in colony is key or the bottleneck of silkworm procreation.During silkworm egg is produced, male moth conventionally can with 2-3 the normal mate and oviposit of female moth, if control male silkworm, support female silkworms more, in the Ci Can colony that makes to raise, shared ratio is by 50% raising 70%, produce the silkworm egg of same quantity, original seed raising scale will dwindle 1/3 in theory, and cost will reduce by 1/3.In addition, raw cocoon production utilizes silkworm sex linked balanced lethal line to breed male silkworm crossbreed in producing, with female-male proportion can not regulate and control, male and female silkworm ratio remains 1:1 the female moth of conventional silkworm kind and the male moth mating production of hybrid seeds of silkworm sex linked balanced lethal line always, breed a male silkworm crossbreed and be a Common corn hybrid production cost of breeding 4 times; If eliminate in the grown silkworm phase the individual hybridization of hero that male individuality is selected and remain female individuality and silkworm sex linked balanced lethal line with speckle sex limited race, the production cost of a male silkworm crossbreed of breeding is 1.75 times an of Common corn hybrid of breeding; If the single female silkworm kind that is 1:0 with female-male proportion is as the female parent of the male silkworm crossbreed of breeding, the production cost of male silkworm crossbreed will significantly reduce, and contribute to social production to accept extensively and raise single male silkworm.
Parthenogenesis is the effective way of getting rid of the single female individuals of male acquisition.Applying many parthenogenesis methods is the lukewarm water facture that the Astaurov of the former Soviet Union proposes, main operation is to take out ovum in the female moth body from sprouting wings, in clear water, remove foreign material, place 10-12 hour in 25 ℃ of environment after, with 46 ℃ of warm water, soak ovum 18min, put into again room temperature water 3-5min, after taking out ovum and drying, under 15-17 ℃, RH80% damp condition, place 3d, can obtain some parthenogenesis and grow ovum.China Wang Yong be take by force bivoltine race as material, and the suitable parthenogenesis lukewarm water inductive condition of thinking current race is to soak 44--46 ℃ of ovum temperature, soak ovum time 14--18 minute.Between parthenogenesis incidence kind, have very big-difference (34.2%-87.6%), average originating rate is 51.13%; The incubation rate that ovum is grown in parthenogenesis is quite low, and some kinds only have 1 or 2 larva hatchings.By the directive breeding of more than 10 years, the parthenogenesis incidence of some strain can reach 90%, and the incubation rate that some strain is grown ovum can reach 80%.The common lopsided silkworm of larva that parthenogenesis hatches, health is poor, difficulty is foster.We raise the individual and female individuality of sexual reproduction of the parthenogenesis of same kind, result parthenogenesis individuality have grow slow, through long, cocoon weight and cocoon shell weight is low, health is poor, difficulty is foster shortcoming.The female silkworm application that these explanation parthenogenesis obtain is produced and is really had many difficulties.
The red ant sch of silkworm sex-linkage (1-21.5) gene pairs Normal appearances is recessive inheritance, and heterozygote newly-hatched silkworm body colour shows as conventional black ant, homozygote newly-hatched silkworm body colour shows as red ant mutant character.Huang Junting utilize chromosome engineering by normal silkworm Z chromosome+the sch fragment normal dyeing body fragment of sch gene (corresponding to) swivel base on W chromosome, obtain sex-limited ant colour system system.The genotype of this system hero is Z
schz
sch, two allelomorph isozygoty and show as red ant; Female genotype is Z
schw
+ sch, on W chromosome, there is the dominant normal gene that sch is corresponding, performance dominant gene proterties is normal black ant.On common silkworm Z chromosome, without sch mutator gene, be normal (+sch), male and female newly-hatched silkworm all shows as black ant, and wherein the genotype of female individuality is Z
+ schw, male individual genotype is Z
+ schz
+ sch.
Summary of the invention
Object one of the present invention is to provide the breeding method of the controlled practical silkworm kind of a kind of male ratio.
Object one of the present invention is to realize by following measures:
A breeding method for the practical silkworm kind that male ratio is controlled, (is called for short Z afterwards to carry sch mutator gene on silkworm Z chromosome
sch, the corresponding normal Z that is called for short
+ sch) and W chromosome on had by swivel base+sch gene (be called for short W
+ sch) the sex-limited ant colour system system of co-controlling is as donor parents, (female genotype is Z to the practical silkworm kind of the common ant look of take
+ schw, Biao Xian Wei ?ant, male genotype is Z
+ schz
+ sch, Biao Xian Wei ?ant) as receptor parent, carry out gene transfer, be about to the Z of donor parents
schgene and W
+ schgene transfer is to receptor parent.
The breeding method of the practical silkworm kind that above-mentioned male ratio is controlled, described gene transfer process comprises the following steps: first respectively by Z
schgene and W
+ schgene transfer, to receptor parent, is set up respectively Z
schgene transfer system and W
+ schgene transfer system, then by Z
schgene transfer system and W
+ schz in gene transfer system
schgene and W
+ schgene fusion to one system, completes the Z of donor parents
schgene and W
+ schgene transfer is to receptor parent.As the practical silkworm kind of receptor parent should be grow neat, health is good, silk good, integral economic trait of fine quality better, there is the silkworm kind of one or more advantage.At Z
schgene and W
+ schgene shift respectively and the last whole process merging in, the selection of the economical and practical proterties of silkworm and raising and stablely all adopt conventional breeding method of mulberry silkworm.
Above-mentioned W
+ schw is set up in gene transfer
+ schgene transfer system comprises the following steps: 1. step, carries out W with the male mating of the female and receptor parent of donor parents
+ schthe hybridization of transfer for the first time of gene; Step 2., the offspring who produces by previous step in pupa time the select and remain male mating of female individuality and receptor parent again carry out W
+ schgene transfer hybridization; Repeating step 2. m time, m>=1.So, W
+ schgene is every need shift hybridization once to receptor parent, and the male mating of just selecting and remain female individuality and receptor parent with the offspring that previous step produces in pupa time, until W
+ scheconomic form proterties and the quantitative character of gene transfer system reach realistic scale.2+m+1 step, it is stable that the offspring that 2+m step produces carries out selfing, completes donor parents W
+ schgene is to the transfer of receptor parent.As shown in Figure 1.The W completing
+ schthe female genotype of gene transfer system is Z
+ schw
+ sch, Biao Xian Wei ?ant, male genotype is Z
+ schz
+ sch, Biao Xian Wei ?ant.
Silkworm Z chromosome is sex crisscross inheritance, and sch is to being normally recessive inheritance, and after the red ant food of sch gene pure body mulberry, body colour mutant character fades away, and plays silkworm 2 ages and starts and common black newly-hatched silkworm no significant difference.In gene transfer process, need male and female sex to be combined with newly-hatched silkworm body colour phenotype the Z that selects and remain
schgene.The second, for gene transfer system obtains more polyceptor parental cell cytoplasmic inheritance material, improve fast its economic achievement, complete Z
schgene transfer, at Z
schin gene transfer process, use as far as possible the female hybridization of receptor parent.Based on these two principles, above-mentioned Z
schz is set up in gene transfer
schgene transfer system comprises the following steps: 1. step, shifts hybridization for the first time with the hero of the female and donor parents of receptor parent; 2. step, shifts that filial generation is female shows as red ant for the first time, and hero shows as black ant, collect ants while raising, eliminate the female Xuan Liu of red ant ?ant is male shifts for the second time hybridization with receptor parent female; 3., shift for the second time filial generation hero has 2 kinds of genotype to step, and phenotype is all black ants, and female have 2 kinds of genotype, and phenotype is divided 2 kinds of the black ants of red ant, collect ants Tao Tai while raising ?the select and remain hero of red ant and receptor parent of ant shift for the third time hybridization; 4. step, shifts for the third time offspring's male and female of hybridization and only has respectively a kind of genotype and a kind of black ant phenotype, and select and remain pupa time male individual and receptor parent female carries out the 4th time and shift hybridization cocoon.4. 3. step form a Z with step
schthe operation cycle of gene transfer hybridization, so circulation is inferior through n (n>=0), until Z
scheconomic form proterties and the quantitative character of gene transfer system reach realistic scale.4+2n+1 step, offspring's hero that 4+2n step produces has 2 kinds of genotype, phenotype is all black ants, female have 2 kinds of genotype, phenotype is divided 2 kinds of the black ants of red ant, collects ants and while raising, the black ant of red ant is separately raised, and red ant is female individuality entirely, in black ant, male and female individuality all has, female hero and the female selfing subculture of red ant of selecting and remain in black ant individuality in cocoon superseded black ant individuality in pupa time; 4+2n+2 step, the offspring that 4+2n+1 step produces has 2 kind of groups, and a types of populations is that in a moth ovum, male and female all only have a kind of genotype, all only has the black ant of a kind of phenotype, this kind of groups does not have target gene type and phenotype, during hatching, according to the phenotype of newly-hatched silkworm, eliminates this kind of groups; Another kind of colony is the interior female 2 kinds of phenotypes of 2 kinds of black ants of the red ant of genotype that have of a moth ovum, and hero also has 2 kinds of phenotypes of 2 kinds of black ants of the red ant of genotype, and red ant hero and red ant are female is Z
schthe target individual of gene transfer demand, eliminates the black ant red ant of selecting and remain and raises, selfing subculture during hatching; 4+2n+3 step, offspring's male and female that 4+2n+2 step produces are all red ants, selfing is stable.So far, complete donor parents Z
schgene is to the transfer of receptor parent.As shown in Figure 2.The Z completing
schthe female genotype of gene transfer system is Z
schw, shows as red ant, and male genotype is Z
schz
sch, also show as red ant.
Above-mentioned W
+ schgene transfer system and Z
schthe fusion of gene transfer system comprises the following steps: 1. step, uses W
+ schfemale (black ant) and the Z of gene transfer system
schthe hero hybridization of gene transfer system; 2., the female genotype of offspring that 1. step produces is Z to step
schw
+ sch, phenotype is black ant, male genotype is Z
schz
+ sch, phenotype is black ant, eliminate in cocoon pupa time hero select and remain female again with Z
schthe hero hybridization of gene transfer system; 3., the female genotype of offspring that 2. step produces is Z to step
schw
+ sch, phenotype is black ant, male genotype is Z
schz
sch, phenotype is red ant, selfing is stable, completes W
+ schgene and Z
schgene fusion to one system.As shown in Figure 3.This is a new sex-limited ant colour system system, and hero has temperature sensitive lethal embryonic period, embryonic phase, and femalely there is no temperature sensitive lethal embryonic period, embryonic phase.So far, complete the Z of donor parents
schgene and W
+ schgene transfer, to receptor parent, obtains the controlled practical silkworm kind of male ratio.
In gene transfer process, gene transfer hybridization is 1 time in theory, and the genetic material of receptor parent accounts for 50% in gene transfer system, gene transfer hybridization 2 times, and the genetic material of receptor parent accounts for 75% in gene transfer system; Hybridize 3 times, the genetic material of receptor parent accounts for 87.5% in gene transfer system; The like, to hybridize n time, the genetic material of receptor parent accounts for [1-(1/2) in gene transfer system
n)] * 100%.If gene transfer hybridization 4～5 times, the genetic material of receptor parent accounts for 93.75～96.875% in gene transfer system, and the genetic material of donor parents only accounts for 3.125～6.25% in gene transfer system.The more, the genetic material of receptor parent shared ratio in gene transfer system is larger for gene transfer hybridization number of times, but hybridization number of times is more, needs from generation to generation manyly, must extend breeding cycle or increase breeding cost.In fact in filial generation, containing the individual binomial probability of obeying of receptor parent blood relationship composition distributes, hybridization number of times is few, and the probability of the defect individual of selecting and remain is less, and hybridization number of times is more, the probability of defect individual selected and remain is larger, contains how many excellent genes keys and be to select in the offspring that reserves seed for planting.At certain experimental size or under the superseded dynamics of certain selection, to hybridize 3～4 times, the defect individual of selecting and remain is feasible.Whether gene transfer continues hybridization after hybridizing 3～4 times, can determine according to the Economic Characters of gene transfer system.
The controlled practical silkworm kind of above-mentioned male ratio is hastened the hatching of silkworms with conventional environment, male and female newly-hatched silkworm hatching ratio (?the red ant hatching of ant ratio) be approximately 1:1.
The object of the invention two is to provide the method that adopts the controlled male ratio of practical silkworm variety control of above-mentioned male ratio.
The object of the invention two realizes by following measures:
Adopt the method for the controlled male ratio of practical silkworm variety control of above-mentioned male ratio, in conventional Incubation environment, hasten the hatching of silkworms to oneself
2～oneself
3 +during embryo, with the humidity environment of 28 ℃～34 ℃ temperature, RH50%～60%, process 24-60 hour, then in extremely hatching of conventional Incubation environment protection.It is 1:1 balance that this method is broken the male and female silkworm ratio of hatching.If wish, obtain the colony that female-male proportion is 2:1～3:1, just by the low-intensity of controlling male ratio, process; If wish, obtain the colony that female-male proportion approaches 1:0, just adopt the high strength of controlling male ratio to process.The processing intensity that gives the different male ratios of control, male and female are hatched ratio difference, can realize the newly-hatched silkworm female-male proportion hatching and be all kinds of female-male proportion colony that 2:1 or 3:1 even approach 1:0.Can freely control male and female silkworm ratio like this, meet multiple needs.
Beneficial effect of the present invention
The present invention is (to be Z by the key gene of controlling male ratio
schgene and W
+ schgene) genetic development and silkworm breeding classical way organically combine, and build the method that forms the controlled practical silkworm kind of the male ratio of a kind of quick breeding.
Advantage of the present invention:
1. can obtain the controlled silkworm kind of male ratio that reaches production application requirement.Original sex-limited ant colour system system is a genetic operating system, and economic characters are very poor, can not directly apply to production at all.The Z that very poor, the very wild original sex-limited ant colour system of economic characters can be united with the present invention
schgene and W
+ schgene transfer is to practical silkworm kind (receptor parent), by practical silkworm kind, the genetic constitution of original sex-limited ant colour system system is replaced and transformed in other words, obtain economic form proterties (as cocoon shape, cocoon look etc.) and quantitative character (as cocoon weight, cocoon shell weight etc.) and approach practical parent, reach realistic scale, can meet the controlled practical silkworm kind of male ratio that silkworm industry is produced a plurality of project checks of a plurality of indexs and requirement.
2. can obtain the controlled practical silkworm kind of the male ratio of multiclass with different characteristics.According to difference, raise season the requirement in (as the spring is used silkworm kind, Xia Qiu silkworm kind etc.) and raising region (as Silkworm region in the north China, southern silkworm district), selection has the practical silkworm kind of corresponding feature and makes receptor parent, with the present invention, can obtain and adapt to the different controlled practical silkworm kinds of the male ratio of multiclass of raising season and raising region.
3. can be to the utilization of the controlled practical silkworm kind advantage of male ratio and the transformation of shortcoming.Can be by the combination of advantages of the controlled silkworm kind of certain male ratio to another good common silkworm variety with the present invention, obtain higher level, the controlled silkworm kind of male ratio that production capacity is stronger; Can be for certain shortcoming of the controlled silkworm kind of male ratio with the present invention, with the common silkworm variety with attendant advantages, transforms, obtain all good controlled practical silkworm kinds of male ratio of each characteristic index.
4. grow normal, health is good, without lopsided silkworm, (parthenogenesis has lopsided silkworm to occur, difficult supporting) occurs.
5. easy and simple to handle, incidence is high.By the controlled practical silkworm kind of male ratio that the present invention obtains, adopt conventional male and female moth mating pattern breeding of future generation, the silkworm seed of the female moth output of post-coitum is nearly all fertilized egg, under each fertilized egg normal condition, can develop into body one by one, easy and simple to handle, incidence is high, almost 100% (parthenogenesis is to take out silkworm seed from virgin moth abdomen, give again specific condition and become two times of growth individualities by it, complex operation, average 50% left and right of incidence, after 10 generation successive selection, some kind incidence can reach 90%, and incidence is low.)。
6. incubation rate is high.The controlled practical silkworm kind of male ratio obtaining with the present invention, under conventional incubation condition, male and female silkworm incubation rate is all more than 90%.By the method for the male ratio of control provided by the invention, control male hatching, can guarantee that the incubation rate of female silkworm is more than 90%, incubation rate is high, (incubation rate of parthenogenesis growth ovum is quite low to reach production real requirement, some kinds only have 1 or 2 larva hatchings, and after the directive breeding of more than 10 year, the incubation rate of some breed development ovum can reach 80%.)。
7. arbitrarily control male and female and hatch ratio, meet multiple demand.The controlled practical silkworm kind of male ratio obtaining with the present invention, sch gene pure body (is Z
schz
schhomozygote hero and Z
schw homozygote is female) red ant has temperature sensitive lethal feature embryonic period, embryonic phase, and the red ant of sex-limited ant colour system system is male, and (genotype is Z
schz
sch) have embryonic period, embryonic phase temperature sensitive lethal ， Er ?female (the genotype Z of ant
schw
+ sch) there is no temperature sensitive lethal an embryonic period, embryonic phase.Under conventional incubation condition, hasten the hatching of silkworms, the male and female silkworm ratio of hatching is approximately 1:1.With the controlled temperature range of male ratio provided by the invention, time range and humidity range, process, guaranteeing that female hatching reaches under the condition of realistic scale, select temperature strength and time intensity can arbitrarily control the male ratio of hatching of the controlled practical silkworm kind of male ratio, freely regulate and control male and female silkworm ratio, meet multiple demand (parthenogenesis only has female silkworm).
Accompanying drawing explanation
The dry tool gene transfer of Fig. 1 W+s flow chart; Individual for eliminating genotype in wire frame; * representative hybridization, ⊕ represents selfing.
Fig. 2 Z
schgene transfer flow chart; Individual for eliminating genotype in wire frame; * representative hybridization, ⊕ represents selfing.
Fig. 3 W
+ schgene transfer system and Z
schgene transfer system merges flow chart; Individual for eliminating genotype in wire frame; * representative hybridization, ⊕ represents selfing.
The controlled practical silkworm kind cyanines pine W of the male ratio of Fig. 4
+ sch, 871W
+ schand the photo of parent's cocoon.
The controlled practical silkworm kind cyanines pine W of the male ratio of Fig. 5
+ sch, 871W
+ schand the photo of parent's ovum.
Fig. 6 controls male percentage effect and represents photo; Red arrow indication male (red ant); A is that low-intensity is controlled effect, and B is that high strength is controlled effect.
embodiment
Below by embodiment, the present invention is specifically described; be necessary to be pointed out that at this; following examples are only used to further illustrate the present invention; can not be interpreted as limiting the scope of the invention, these those skilled in the art can make some nonessential improvement and adjustment to the present invention according to the invention described above content.
The sex-limited ant colour system system Silkworm Variety Resource W-01 that adopts Biotechnology Institute of Southwestern University to preserve, this system is a genetic operating system, economic characters very poor (specifically in Table 1, Fig. 4 and Fig. 5), can not be applied to produce.We make donor parents with W-01, be used in produce large-scale popularization application the practical silkworm kind of high-quality Chinese system cyanines pine, 871 etc. as receptor parent, utilize the flow process of Fig. 1 (m=2), Fig. 2 (n=1), Fig. 3, by the Z of W-01 system
schgene and W
+ schgene proceeds to wherein, obtains the controlled cyanines pine silkworm kind of male ratio and (is called for short cyanines pine W
+ sch) and controlled 871 silkworm kinds (the abbreviation 871W of male ratio
+ sch).
A. the practical effect of economic characters
Compare with original sex-limited ant colour system system W-01, the controlled practical silkworm breed development of male ratio omits slow, but grow neat, economic form proterties homogeneous, the cocoon shape and the ovum look feature (seeing Fig. 4 and Fig. 5) that have obvious Chinese system receptor parent, economic quantities proterties is greatly improved, and approaches with receptor parent, reaches realistic scale (in Table 1).With 5 length of times, through growing process with full age through investigating, with worm pupa rate investigation health, with single moth egg amount, investigate and breed proterties, with cocoon weight, cocoon shell weight and cocoon layer rate, investigate Cocoon Quality Traits, the controlled practical silkworm kind cyanines pine W of male ratio
+ schand 871W
+ sch, corresponding receptor parent cyanines pine and 871 and donor parents W-01, growth process, the health of raising autumn in 2013, breed achievement and cocoon quality achievement as table 1.As known from Table 1, though the controlled practical silkworm breed development of male ratio is slightly slower than donor parents, faster than receptor parent, and at health, breed the aspects such as performance and cocoon quality achievement far beyond donor parents, approach with receptor parent.From Fig. 4 and Fig. 5, the controlled practical silkworm kind economic form proterties of male ratio is as cocoon shape, cocoon body size, ovum look and produce attached etc. similarly to receptor parent, and economic form is neat and well spaced good, meets production application requirement.
Table 1 main rearing autumn in 2013 (single moth ovum number, cocoon quality achievement are 10 individual average achievements of sampling)
B. the controlled effect of male ratio
Autumn in 2013, by the instant pickling kind of the controlled silkworm kind of male ratio obtaining, is controlled the specially treated of male ratio in embryo development procedure, obtains corresponding male and female newly-hatched silkworm and hatches ratio.
1. control the effect that female-male proportion is 2:1--3:1
When embryo is developed to oneself in conventional Incubation environment
2 +or oneself
3period, with 28 ℃, the condition of RH60%, process 24 hours, and then in conventional Incubation environment protection to hatching.Hatching investigation result is as table 2.871W
+ schvarietal Investigation 2153, wherein black ant is female has hatched 1058, red ant hero has been hatched 401, the red ant ratio of black ant is that female-male proportion is about 2.6:1, meeting seed station, to breed original seed female than male many ratio needs; By male and female in total ovum grain number, be 1:1, the female incubation rate of black ant is 98.26%, arrives Practical Hatchability of Silkworm Eggs required standard more than 88%.Cyanines pine W
+ schkind has 3972 ovum, and black ant is female has hatched 1923, and the female incubation rate of black ant is 96.87%, and red ant hero has been hatched 631, and the male and female ratio of hatching is about 3:1, male rate control effect and realistic scale and 871W
+ schkind is similar.Hatching represents that photo is shown in Fig. 6.
It is 2:1--3:1 experimental investigation result that the male ratio of table 2 control makes female-male proportion
2. control the male effect that makes the approximate single female silkworm of colony
When embryo is developed to oneself in conventional Incubation environment
2 +or oneself
3period, with 32 ℃, the condition of RH60%, process 48 hours, then in conventional Incubation environment protection to hatching.Hatching investigation result is as table 3.At 2402 871W
+ schsilkworm seed in, have the male hatching of the red ant of 1116 female hatchings of black ant, 57, the female incubation rate of black ant is 96.87%, in newly-hatched silkworm, the female ratio of black ant is 95.14%; 1857 cyanines pine W that process
+ schin silkworm seed, black ant is female has hatched 873, and the female incubation rate of black ant is 92.92%, and red ant hero has been hatched 10, and in newly-hatched silkworm, the female ratio of black ant is 98.97%.Two female incubation rates of kind all reach realistic scale, and female silkworm ratio is all more than 95%, can be described as single female silkworm.Fig. 6 is shown in hatching representative.
Table 3 is controlled the experimental investigation result that male ratio makes the approximate single female silkworm of colony
Illustrate: 871W
+ schwith cyanines pine W
+ schtwo kinds, except newly-hatched silkworm body colour is subject to homologous genes controls, other genetic background widely different (the 871st, spring and autumn dual-purpose kind, and cyanines pine is to use silkworm kind the spring.), so they are slightly variant to the treatment conditions reaction of male rate control.
C. the controlled silkworm kind of male ratio obtaining is grown normally, and health is good, without lopsided silkworm, occurs; Easy and simple to handle, incidence is high, reaches 100%; Incubation rate is high, reaches more than 90%, reaches production real requirement.
Claims (4)
1. a breeding method for the controlled practical silkworm kind of male ratio, to carry on silkworm Z chromosome
schmutator gene (is called for short Z afterwards
sch , the corresponding normal Z that is called for short
+
sch ) and W chromosome on by swivel base, had+
schgene (is called for short W
+
sch ) the sex-limited ant colour system system of co-controlling is as donor parents, (female genotype is Z to the practical silkworm kind of the common ant look of take
+
sch w, Biao Xian Wei ?ant, male genotype is Z
+
sch z
+
sch , Biao Xian Wei ?ant) as receptor parent, carry out gene transfer, by the Z of donor parents
sch gene and W
+
sch gene transfer is to receptor parent.
2. the breeding method of the controlled practical silkworm kind of male ratio as claimed in claim 1, described gene transfer comprises the following steps: first respectively by Z
sch gene and W
+
sch gene transfer, to receptor parent, is set up respectively Z
sch gene transfer system and W
+
sch gene transfer system, then by Z
sch gene transfer system and W
+
sch z in gene transfer system
sch gene and W
+
sch gene fusion to one system, completes the Z of donor parents
sch gene and W
+
sch gene transfer is to receptor parent.
3. the breeding method of the controlled practical silkworm kind of male ratio as claimed in claim 2, described W
+
sch w is set up in gene transfer
+
sch gene transfer system comprises the following steps: 1. step, carries out W with the male mating of the female and receptor parent of donor parents
+
sch the hybridization of transfer for the first time of gene; Step 2., the offspring who produces by previous step in pupa time the select and remain male mating of female individuality and receptor parent carry out W
+
sch the hybridization of transfer again of gene; Repeating step 2. m time, m≤0, the W completing
+
sch the female genotype of gene transfer system is Z
+
sch w
+
sch , Biao Xian Wei ?ant, male genotype is Z
+
sch z
+
sch , Biao Xian Wei ?ant;
Described Z
sch z is set up in gene transfer
sch gene transfer system comprises the following steps: 1. step, shifts hybridization for the first time with the hero of the female and donor parents of receptor parent; 2. step, shifts that filial generation is female shows as red ant for the first time, and hero shows as black ant, collect ants while raising, eliminate the female Xuan Liu of red ant ?ant is male shifts for the second time hybridization with receptor parent female; 3., shift for the second time filial generation hero has 2 kinds of genotype to step, and phenotype is all black ants, and female have 2 kinds of genotype, and phenotype is divided 2 kinds of the black ants of red ant, collect ants Tao Tai while raising ?the select and remain hero of red ant and receptor parent of ant shift for the third time hybridization; 4. step, shifts for the third time offspring's male and female of hybridization and only has respectively a kind of genotype and a kind of black ant phenotype, and select and remain pupa time male individual and receptor parent female carries out the 4th time and shift hybridization cocoon; 4. 3. step form a Z with step
sch the operation cycle of gene transfer hybridization, so circulation is through n(n≤0) inferior, until Z
sch economic form proterties and the quantitative character of gene transfer system reach realistic scale; 4+2n+1 step, offspring's hero that 4+2n step produces has 2 kinds of genotype, phenotype is all black ants, female have 2 kinds of genotype, phenotype is divided 2 kinds of the black ants of red ant, collects ants and while raising, the black ant of red ant is separately raised, and red ant is female individuality entirely, in black ant, male and female individuality all has, female hero and the female selfing subculture of red ant of selecting and remain in black ant individuality in cocoon superseded black ant individuality in pupa time; 4+2n+2 step, the offspring that 4+2n+1 step produces has 2 kind of groups, and a types of populations is that in a moth ovum, male and female all only have a kind of genotype, all only has the black ant of a kind of phenotype, this kind of groups does not have target gene type and phenotype, during hatching, according to the phenotype of newly-hatched silkworm, eliminates this kind of groups; Another kind of colony is the interior female 2 kinds of phenotypes of 2 kinds of black ants of the red ant of genotype that have of a moth ovum, and hero also has 2 kinds of phenotypes of 2 kinds of black ants of the red ant of genotype, and red ant hero and red ant are female is Z
sch the target individual of gene transfer demand, eliminates the black ant red ant of selecting and remain and raises, selfing subculture during hatching; 4+2n+3 step, offspring's male and female that 4+2n+2 step produces are all red ants, selfing is stable; So far, complete donor parents Z
sch gene is to the transfer of receptor parent, the Z completing
sch the female genotype of gene transfer system is Z
sch w, shows as red ant, and male genotype is Z
sch z
sch , also show as red ant;
Described W
+
sch gene transfer system and Z
sch the fusion of gene transfer system comprises the following steps: 1. step, uses W
+
sch female and the Z of gene transfer system
sch the hero hybridization of gene transfer system; 2., the female genotype of offspring that 1. step produces is Z to step
sch w
+ sch, phenotype is black ant, male genotype is Z
sch z
+
sch , phenotype is black ant, eliminate in cocoon pupa time hero select and remain female again with Z
sch the hero hybridization of gene transfer system; 3., the female genotype of offspring that 2. step produces is Z to step
sch w
+
sch , phenotype is black ant, male genotype is Z
sch z
sch , phenotype is red ant, selfing is stable, completes W
+
sch gene and Z
sch gene fusion to one system.
4. adopt the method for the controlled male ratio of practical silkworm variety control of the arbitrary described male ratio of claim 1-3, in conventional Incubation environment, hasten the hatching of silkworms to oneself
2～oneself
3 +during embryo, with the humidity environment of 28 ℃～34 ℃ temperature, RH50%～60%, process 24-60 hour, then in extremely hatching of conventional Incubation environment protection.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110352918A (en) * | 2019-08-15 | 2019-10-22 | 重庆三峡农业科学院(重庆市万州区甘宁蚕种场) | A kind of raising quality of silkworm eggs method |
| CN114216870A (en) * | 2021-10-29 | 2022-03-22 | 辽宁省蚕业科学研究所 | Screening method and application of tussah breeding material |
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| CN1419821A (en) * | 2001-11-20 | 2003-05-28 | 成都天友生物科技股份有限公司 | Temp-sensitive lethality male silkworm breeding method |
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| CN1419821A (en) * | 2001-11-20 | 2003-05-28 | 成都天友生物科技股份有限公司 | Temp-sensitive lethality male silkworm breeding method |
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| 陈萍: "家蚕胚胎期伴性温敏性与伴性赤蚁sch基因及亲本的关系研究", 《蚕业科学》, vol. 33, no. 2, 30 June 2007 (2007-06-30) * |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110352918A (en) * | 2019-08-15 | 2019-10-22 | 重庆三峡农业科学院(重庆市万州区甘宁蚕种场) | A kind of raising quality of silkworm eggs method |
| CN114216870A (en) * | 2021-10-29 | 2022-03-22 | 辽宁省蚕业科学研究所 | Screening method and application of tussah breeding material |
| CN114216870B (en) * | 2021-10-29 | 2023-10-03 | 辽宁省蚕业科学研究所 | Screening method of tussah breeding material and application thereof |
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