CN107646672A - A kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness - Google Patents
A kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness Download PDFInfo
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Abstract
The invention discloses a kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness, comprise the following steps：B and A hybridizes to obtain F1；F1It is selfed to obtain F2, then with A be returned, obtain BC1F1；BC1F1It is selfed to obtain BC1F2, then with A be returned, obtain BC2F1；BC2F1It is selfed to obtain BC2F2, then with A be returned, obtain BC3F1；BC3F1It is selfed to obtain BC3F2, then with A be returned, obtain BC4F1；BC4F1It is selfed to obtain BC3F2, then with A be returned, obtain BC5F1；BC4F1It is selfed to obtain BC5F2, then with A be returned, obtain BC6F1；BC6F1It is selfed to obtain BC6F2, then it is selfed to obtain BC6F3；Above-mentioned self-mating system again with other hybridizations between selfed lines.The advantage of the invention is that：It is simple to provide a kind of step, it is easy to operate, the self-mating system breeding method of the anti-epidemic disease evil Dendrobidium huoshanness for the Dendrobidium huoshanness for taking into account anti-epidemic disease and excellent economical character can be obtained.
Description
Technical field
The present invention relates to Dendrobidium huoshanness to cultivate field, more particularly to a kind of self-mating system cultivation side of anti-epidemic disease evil Dendrobidium huoshanness
Method.
Background technology
Huoshan is located in Anhui Province western part, foot of Dabie Mountains, positioned at 03 '~31 ° 33 ' of north latitude 31 °, 115 ° 52 ' of east longitude
Between~116 ° 32 ', the zone of transition of monsoon climate and temperate zone semi-moist monsoon climate is moistened in subtropical zone, is the orchid family stem of noble dendrobium
Platymiscium geographical distribution is northernmost.Huoshan and its neighbouring mountain area three kinds of medicinal dendrobiums of main product, are declared jointly by Lu'an City government
For geography symbol product, its kind of name is respectively：Dendrobidium huoshanness (Dendrobium huoshanense C.Z.Tang et
S.J.Cheng), also known as meter Xin Shi dry measures used in former times, dendrobium candidum (Dendrobium candidum Wall.exLindl., also known as black section
Grass), and dendrobium moniliformeSweet (Dendrobium moniliforme (Linn.) Sw., also known as Dendrobium Moniliforme), three is orchid family stone
Dry measure used in former times category herbaceos perennial, the active polysaccharide composition mainly contained, there is antitumor, immunological regulation, anti-oxidant, expansion blood
The multiple efficacies such as pipe, hypoglycemic.
The disease that stem of noble dendrobium epidemic disease occurs in hot and humid environment, germ diseased plant or with egg spore etc. in invalid body and soil
Survived the winter in earth；Propagation is splashed by wind and rain or water droplet drop, expection of diving is short, 3-5 days or so.The disease occurs sooner or later sooner or later and secondary with rainfall
Several, warm indoor air relative humidity is closely related, susceptible disease and more serious during rainy, high humidity；Water spray, ventilation difference aggravate disease
Occur；Temperature is less than 25 DEG C, easily popular disease of being cleared up after continuous cloudy day or wet weather.The disease often makes plant young tender part rot, funeral
Lose commodity value；The disease makes plant top lobus cardiacus base portion that water stain shape brown occur to rot, and to leaf tip extension；The disease causes stem
In water stain shape brown rot, stem is set all to rot when seriously ill, and spread speed is fast, infection area is big.
Self-mating system refers to an individual plant after continuous selfing, the offspring of the character neat and consistent obtained.Cross-pollination is made
Thing, under conditions of general field planting, between different cultivars (being), different plants, easy a string of flowers (powder), more than 95% seed
Grain, all it is solid by the pollination of other plant pollen.If the pollen of this plant is awarded on the pistil stigma of homophyletic, it is allowed to be fertilized
Solid, this just cries selfing.The seed sowing of Second Year selfing, therefrom selects fine individual plant to continue to be selfed, and so continuous selfing 4~
After 5 generations, breeding time, seedling color, plant type, blade profile, plant height, fringe position and Ear Characters of its offspring etc. all can neat and consistent, become for
One self-mating system.
Cross pollinated plant, natural hybrization rate is high, and each kind is essentially all one and mixes colony, and heredity is very not
It is pure.With the impure parents of such heredity, not only the bad character of parents can entail cenospecies, and advantage is low, and F1
In generation, occurs trait segregation and grows irregular.In in general crop varieties colony, the recessive gene for restricting some characters is normal
Covered by simultaneous corresponding dominant allele, so recessive character is not generally shown, so during seed selection cenospecies,
Typically without ready-made kind with self-mating system come preparing hybrid kind.By artificial autocopulation, trait segregation can occur for offspring, many
Bad recessive character is showed, and white seedling, flower seedling, deformity, leaf roll, narrow leaf, small ear, infertility, male flower is such as occurred and is set seeds, easily
It is susceptible, lodging etc., by selection, eliminate it is bad stay it is excellent.So, its gene is understood more and more homozygous and turns into homozygote, and offspring is not
Separate again.So self-mating system can not only overcome the shortcomings that original kind, its merit is set to be accumulated, strengthened；It is and each
The character of individual plant all very stable and consistents.Be hybridized with the different self-mating system of heredity simultaneously after, the viability of F1 generation can assemble respectively
The merit of parental inbred line, larger hybrid vigour is showed, also neat and consistent, its economical character are better than kind or product to character
Interspecific hybrid.General volume increase 20~30%.
Back cross breeding method is that the improved seeds for lacking a certain two beneficial traits are reused as into parent, claims samsara parent
This, and because being the recipient of beneficial traits, also referred to as acceptor.Recurrent parent should be that Comprehensive Traits are preferable, it is contemplated that promising
Kind.With another there is the kind of beneficial traits lacking in a certain two acceptors to make parent, only when starting backcrossing with 1 time,
Claim nonrecurrent parent.Because being the supplier of beneficial traits, therefore claim donor.In backcross process, since first backcross generation, per Dai Doucong
The individual of beneficial traits of the selection with donor hybridizes with recurrent parent in hybrid.So continue repeatedly, to the last
It is similar to acceptor to all characters, but when adding the offspring of the beneficial traits transferred from donor untill.Then 1~2 is carried out again
Secondary selfing, select and be transferred character as homozygous individual, and then be bred as new varieties.Often it is returned 1 time in theory, hybrid generation institute
Genetic constitution containing recurrent parent will be incremented by half.Typically through 5~6 backcrossings, the major traits of its offspring are close to samsara parent
This.But the gene number that the major traits such as recurrent parent are related to is more, then being returned number will suitably increase.
The program of back cross breeding
1. hybridization hybridizes the recurrent parent and nonrecurrent parent selected.
2. backcrossing is selected the plant with objective trait from backcross progeny and is returned with recurrent parent.
3. selfing is after being returned and arriving the suitable generation, it is necessary to is selfed the generation of l~2.Because the scape character of backcross progeny at this moment
It is substantially homozygous, but objective trait is still within heterozygous state, it is necessary to be selfed purifying, could obtain the stable strain of a character.
The method of existing Breeding of Inbred Lines mainly using the method for two rings choosing system, i.e., is sent to certainly after donor and acceptor hybridization
Bottom, the final self-mating system for obtaining merit.However, existing utilize the method that the choosing of two rings is seed selection merit self-mating system, by
In being typically all to use 2 hybridizations between selfed lines, offspring's self cross and selection, easily cause breeding material hereditary basis narrow, subsequently educate
Into kind have and high feel certain disease and cause the risk lost greatly.And the related gene of the excellent kind of economical character often with
Some bad character gene (such as susceptible gene) linkage inheritances.Therefore, there is an urgent need to provide a kind of new seed selection selfing in the industry
The method of system, to overcome the problems of prior art.
The content of the invention
It is easy to operate it is an object of the invention to overcome the deficiencies of the prior art and provide a kind of step is simple, it can obtain
Take into account the self-mating system breeding method of the anti-epidemic disease evil Dendrobidium huoshanness of the Dendrobidium huoshanness of anti-epidemic disease and excellent economical character.
The present invention is achieved by the following technical solutions：A kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness,
Comprise the following steps：
(1) antilemic plant system B and economical character excellent self-mating system A is hybridized, obtains hybridizing F1；
(2) the excellent F of economical character is chosen1Individual plant selfing, obtain selfing F2, in seedling stage using epidemic disease in vitro with base of leaf bar
Identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC1F1；
(3) the excellent BC of economical character is chosen1F1Individual plant selfing, obtain selfing BC1F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC2F1；
(4) the excellent BC of economical character is chosen2F1Individual plant selfing, obtain selfing BC2F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC3F1；
(5) the excellent BC of economical character is chosen3F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC4F1；
(6) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC5F1；
(7) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC5F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC6F1；
(8) the excellent BC of economical character is chosen6F1Individual plant selfing, obtain selfing BC6F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then individual plant selfing, obtains BC6F3, anti-epidemic disease is shown as from explanation, finally
Obtain high anti-epidemic disease and the excellent self-mating system of economical character；
(9) the high anti-epidemic disease obtained by above-mentioned steps and the excellent self-mating system of economical character are carried out with other self-mating systems again miscellaneous
Hand over, high antilemic Dendrobidium huoshanness cenospecies can be selected.
One of preferred embodiment as the present invention, the excellent finger plant of economical character have high yield, high-quality, good stress resistance
Breeding objective in one or more of states.
As one of preferred embodiment of the present invention, the excellent Dendrobidium huoshanness seedling of the economical character of the selection carry out selfing and
During backcrossing, the strain number of selection accounts for the 10-15% of total strain number.
One of preferred embodiment as the present invention, the total strain number are no less than 50 plants.
One of preferred embodiment as the present invention, band base of leaf bar identification method is specially the epidemic disease in vitro：Taken in seedling stage together
Tether panel base of leaf bar, 8-12min is rinsed with clear water, with 2-4% hypochlorite disinfectant 6-10min, finally rushes Xian with sterilized water
3-5 times, being covered with the culture dish of double-deck moistening filter paper for sterilization is face-up put into, it is (0.5- that surface, which sprays into concentration,
1.5)×105Individual/ml epidemic disease spore suspensions, alternation of light and darkness in 22 DEG C/12h, 18 DEG C/12h incubator is placed in after sealing and is trained
2-4d is supported, then carries out anti-epidemic disease identification and the statistical analysis of disease index.
One of preferred embodiment as the present invention, the disease scale standard that the anti-epidemic disease is identified is：It is 0 grade, asymptomatic；1
Grade, there is a small amount of epidemic disease shape scab on blade and scab is smudgy, lesion area accounts for less than the 5% of the gross area；2 grades, on blade
There is obvious scab, but it is scattered not in flakes, lesion area accounts for the 6%-25% of the gross area；3 grades, moderate quatity is produced on blade
Epidemic disease shape scab and scab are linked to be piece, and lesion area accounts for the 26%-50% of the gross area；4 grades, the epidemic disease Lesion number of blade is more,
Account for the 51%-75% of leaf area；5 grades, the epidemic disease Lesion number on blade is many and bisque is thicker, accounts for the 76%- of leaf area
100%；
The statistical analysis technique of the disease index is：Disease index mirror is calculated using the statistical analysis formula of disease index
Fixed, standard of perfection is：Height is anti-, 0 ＜ disease index≤15；It is disease-resistant, 15 ＜ disease index≤35；Moderate resistance, 35 ＜ disease indexs≤
55；It is susceptible, 55 ＜ disease index≤75；Height sense, disease index >=75.
One of preferred embodiment as the present invention, the statistical analysis formula of the disease index is：DI=Σ (s × n) ×
100/N × S, wherein, DI represents disease index, and s represents the representative numerical value of each state of an illness rank, and n represents each state of an illness rank disease number of sheets,
N represents investigation total number of sheets, and S represents highest level typical value.
One of preferred embodiment as the present invention, other self-mating systems are specially to have high anti-southern blight in the step (9)
Self-mating system, the self-mating system with high polysaccharide or the self-mating system with high anti-anthracnose.
One of preferred embodiment as the present invention, selects whole tissue culture.
The present invention compared with prior art the advantages of be：
(1) seedling stage using Dendrobidium huoshanness in vitro with base of leaf bar to backcross progeny carry out epidemic disease identification and screening, have
It is easy, efficiently, the advantages of accuracy rate is high；
(2) the high anti-epidemic disease kind that the present invention is bred as, possesses the characteristics of disease resistance is strong and economical character is good, raw in agricultural
Popularization and application in production, it is possible to reduce the use of chemical pesticide, both favorably produced nontoxic green product and met consumption demand,
Preserve the ecological environment again and promote the sustainable development of agricultural, there is good economic benefit and social benefit；
(3) after obtaining high anti-epidemic disease and the excellent self-mating system of economical character, then by its with possess other merits from
Hand over system to be hybridized, high antilemic Dendrobidium huoshanness cenospecies can be selected, the cenospecies not only possesses disease resistance by force and agronomy
The characteristics of character is good, it is also equipped with the merit needed for other；
(4) tissue culture is used, cultivation period can be accelerated.
Brief description of the drawings
Fig. 1 is a kind of overall flow step of the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness in embodiment 1-3
Schematic diagram.
Embodiment
Embodiments of the invention are elaborated below, the present embodiment is carried out lower premised on technical solution of the present invention
Implement, give detailed embodiment and specific operating process, but protection scope of the present invention is not limited to following implementation
Example.
Embodiment 1
As shown in figure 1, a kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness of the present embodiment, including following step
Suddenly：
(1) antilemic plant system B and economical character excellent self-mating system A is hybridized, obtains hybridizing F1；
(2) the excellent F of economical character is chosen1Individual plant selfing, obtain selfing F2, in seedling stage using epidemic disease in vitro with base of leaf bar
Identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC1F1；
(3) the excellent BC of economical character is chosen1F1Individual plant selfing, obtain selfing BC1F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC2F1；
(4) the excellent BC of economical character is chosen2F1Individual plant selfing, obtain selfing BC2F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC3F1；
(5) the excellent BC of economical character is chosen3F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC4F1；
(6) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC5F1；
(7) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC5F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC6F1；
(8) the excellent BC of economical character is chosen6F1Individual plant selfing, obtain selfing BC6F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then individual plant selfing, obtains BC6F3, anti-epidemic disease is shown as from explanation, finally
Obtain high anti-epidemic disease and the excellent self-mating system of economical character；
(9) by the high anti-epidemic disease obtained by above-mentioned steps and the excellent self-mating system of economical character again with high anti-southern blight
Self-mating system is hybridized, and can select high anti-epidemic disease and southern blight and the excellent Dendrobidium huoshanness cenospecies of economical character.
Preferably, the excellent finger plant of economical character has one kind in high yield, high-quality, good stress resistance Breeding objective
Or several state.
Preferably, when the excellent Dendrobidium huoshanness seedling of the economical character of selection is selfed and is returned, the strain number of selection accounts for always
The 10% of strain number, total strain number are 50 plants.
Preferably, band base of leaf bar identification method is specially epidemic disease in vitro：Same tether panel base of leaf bar is taken in seedling stage, is rinsed with clear water
8min, with 2% hypochlorite disinfectant 6min, finally rush Xian 3 times with sterilized water, it is double to be face-up put into being covered with for sterilization
In the culture dish of layer moistening filter paper, it is 0.5- × 10 that surface, which sprays into concentration,5Individual/ml epidemic disease spore suspensions, 22 are placed in after sealing
DEG C/12h, 18 DEG C/12h incubator in alternation of light and darkness culture 2d, then carry out the identification of anti-epidemic disease and the statistical analysis of disease index；
The disease scale standard that anti-epidemic disease is identified is：It is 0 grade, asymptomatic；1 grade, occur a small amount of epidemic disease shape scab and disease on blade
Spot is smudgy, and lesion area accounts for less than the 5% of the gross area；2 grades, occur obvious scab on blade, but it is scattered not in flakes, scab
Area accounts for the 6%-25% of the gross area；3 grades, produce the epidemic disease shape scab of moderate quatity on blade and scab is linked to be piece, lesion area
Account for the 26%-50% of the gross area；4 grades, the epidemic disease Lesion number of blade is more, accounts for the 51%-75% of leaf area；5 grades, on blade
Epidemic disease Lesion number it is many and bisque is thicker, account for the 76%-100% of leaf area；
The statistical analysis technique of disease index is：Utilize the statistical analysis formula DI=Σ (s × n) × 100/ of disease index
(DI- disease indexs, the representative numerical value of each state of an illness ranks of s-, each state of an illness rank disease numbers of sheets of n-, N- investigation total number of sheets, S- is most by N × S
High-level representative is worth) disease index identification is calculated, standard of perfection is：Height is anti-, 0 ＜ disease index≤15；Disease-resistant, the 15 ＜ state of an illness refer to
Number≤35；Moderate resistance, 35 ＜ disease index≤55；It is susceptible, 55 ＜ disease index≤75；Height sense, disease index >=75.
Preferably, only needed 6-8 months because tissue culture is bloomed, and cultivating to bloom needs 14-16 months, so in said process
Whole tissue culture is selected, makes the cycle shorter.
Embodiment 2
As shown in figure 1, a kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness of the present embodiment, including following step
Suddenly：
(1) antilemic plant system B and economical character excellent self-mating system A is hybridized, obtains hybridizing F1；
(2) the excellent F of economical character is chosen1Individual plant selfing, obtain selfing F2, in seedling stage using epidemic disease in vitro with base of leaf bar
Identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC1F1；
(3) the excellent BC of economical character is chosen1F1Individual plant selfing, obtain selfing BC1F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC2F1；
(4) the excellent BC of economical character is chosen2F1Individual plant selfing, obtain selfing BC2F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC3F1；
(5) the excellent BC of economical character is chosen3F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC4F1；
(6) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC5F1；
(7) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC5F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC6F1；
(8) the excellent BC of economical character is chosen6F1Individual plant selfing, obtain selfing BC6F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then individual plant selfing, obtains BC6F3, anti-epidemic disease is shown as from explanation, finally
Obtain high anti-epidemic disease and the excellent self-mating system of economical character；
(9) by the high anti-epidemic disease obtained by above-mentioned steps and the excellent self-mating system of economical character again with the selfing with high polysaccharide
System is hybridized, and can select high anti-epidemic disease, high polysaccharide and the excellent Dendrobidium huoshanness cenospecies of economical character.
Preferably, the excellent finger plant of economical character has one kind in high yield, high-quality, good stress resistance Breeding objective
Or several state.
Preferably, when the excellent Dendrobidium huoshanness seedling of the economical character of selection is selfed and is returned, the strain number of selection accounts for always
The 13% of strain number, total strain number are 80 plants.
Preferably, band base of leaf bar identification method is specially epidemic disease in vitro：Same tether panel base of leaf bar is taken in seedling stage, is rinsed with clear water
10min, with 3% hypochlorite disinfectant 8min, finally rush Xian 4 times with sterilized water, be face-up put into being covered with for sterilization
In the culture dish of bilayer moistening filter paper, it is 1.0 × 10 that surface, which sprays into concentration,5Individual/ml epidemic disease spore suspensions, 22 are placed in after sealing
DEG C/12h, 18 DEG C/12h incubator in alternation of light and darkness culture 3d, then carry out the identification of anti-epidemic disease and the statistical analysis of disease index；
The disease scale standard that anti-epidemic disease is identified is：It is 0 grade, asymptomatic；1 grade, occur a small amount of epidemic disease shape scab and disease on blade
Spot is smudgy, and lesion area accounts for less than the 5% of the gross area；2 grades, occur obvious scab on blade, but it is scattered not in flakes, scab
Area accounts for the 6%-25% of the gross area；3 grades, produce the epidemic disease shape scab of moderate quatity on blade and scab is linked to be piece, lesion area
Account for the 26%-50% of the gross area；4 grades, the epidemic disease Lesion number of blade is more, accounts for the 51%-75% of leaf area；5 grades, on blade
Epidemic disease Lesion number it is many and bisque is thicker, account for the 76%-100% of leaf area；
The statistical analysis technique of disease index is：Utilize the statistical analysis formula DI=Σ (s × n) × 100/ of disease index
(DI- disease indexs, the representative numerical value of each state of an illness ranks of s-, each state of an illness rank disease numbers of sheets of n-, N- investigation total number of sheets, S- is most by N × S
High-level representative is worth) disease index identification is calculated, standard of perfection is：Height is anti-, 0 ＜ disease index≤15；Disease-resistant, the 15 ＜ state of an illness refer to
Number≤35；Moderate resistance, 35 ＜ disease index≤55；It is susceptible, 55 ＜ disease index≤75；Height sense, disease index >=75.
Preferably, only needed 6-8 months because tissue culture is bloomed, and cultivating to bloom needs 14-16 months, so in said process
Whole tissue culture is selected, makes the cycle shorter.
Embodiment 3
As shown in figure 1, a kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness of the present embodiment, including following step
Suddenly：
(1) antilemic plant system B and economical character excellent self-mating system A is hybridized, obtains hybridizing F1；
(2) the excellent F of economical character is chosen1Individual plant selfing, obtain selfing F2, in seedling stage using epidemic disease in vitro with base of leaf bar
Identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC1F1；
(3) the excellent BC of economical character is chosen1F1Individual plant selfing, obtain selfing BC1F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC2F1；
(4) the excellent BC of economical character is chosen2F1Individual plant selfing, obtain selfing BC2F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC3F1；
(5) the excellent BC of economical character is chosen3F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC4F1；
(6) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC3F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC5F1；
(7) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC5F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC6F1；
(8) the excellent BC of economical character is chosen6F1Individual plant selfing, obtain selfing BC6F2, it is in vitro using epidemic disease in seedling stage
Band base of leaf bar identification method carries out anti-epidemic disease identification, then individual plant selfing, obtains BC6F3, anti-epidemic disease is shown as from explanation, finally
Obtain high anti-epidemic disease and the excellent self-mating system of economical character；
(9) by the high anti-epidemic disease obtained by above-mentioned steps and the selfing with high anti-anthracnose again of the excellent self-mating system of economical character
System is hybridized, and can select high anti-epidemic disease and anti-anthracnose and the excellent Dendrobidium huoshanness cenospecies of economical character.
Preferably, the excellent finger plant of economical character has one kind in high yield, high-quality, good stress resistance Breeding objective
Or several state.
Preferably, when the excellent Dendrobidium huoshanness seedling of the economical character of selection is selfed and is returned, the strain number of selection accounts for always
The 15% of strain number, total strain number are 100 plants.
Preferably, band base of leaf bar identification method is specially epidemic disease in vitro：Same tether panel base of leaf bar is taken in seedling stage, is rinsed with clear water
12min, with 4% hypochlorite disinfectant 10min, finally rush Xian 5 times with sterilized water, be face-up put into being covered with for sterilization
In the culture dish of bilayer moistening filter paper, it is 1.5 × 10 that surface, which sprays into concentration,5Individual/ml epidemic disease spore suspensions, 22 are placed in after sealing
DEG C/12h, 18 DEG C/12h incubator in alternation of light and darkness culture 4d, then carry out the identification of anti-epidemic disease and the statistical analysis of disease index；
The disease scale standard that anti-epidemic disease is identified is：It is 0 grade, asymptomatic；1 grade, occur a small amount of epidemic disease shape scab and disease on blade
Spot is smudgy, and lesion area accounts for less than the 5% of the gross area；2 grades, occur obvious scab on blade, but it is scattered not in flakes, scab
Area accounts for the 6%-25% of the gross area；3 grades, produce the epidemic disease shape scab of moderate quatity on blade and scab is linked to be piece, lesion area
Account for the 26%-50% of the gross area；4 grades, the epidemic disease Lesion number of blade is more, accounts for the 51%-75% of leaf area；5 grades, on blade
Epidemic disease Lesion number it is many and bisque is thicker, account for the 76%-100% of leaf area；
The statistical analysis technique of disease index is：Utilize the statistical analysis formula DI=Σ (s × n) × 100/ of disease index
(DI- disease indexs, the representative numerical value of each state of an illness ranks of s-, each state of an illness rank disease numbers of sheets of n-, N- investigation total number of sheets, S- is most by N × S
High-level representative is worth) disease index identification is calculated, standard of perfection is：Height is anti-, 0 ＜ disease index≤15；Disease-resistant, the 15 ＜ state of an illness refer to
Number≤35；Moderate resistance, 35 ＜ disease index≤55；It is susceptible, 55 ＜ disease index≤75；Height sense, disease index >=75.
Preferably, only needed 6-8 months because tissue culture is bloomed, and cultivating to bloom needs 14-16 months, so in said process
Whole tissue culture is selected, makes the cycle shorter.
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 be included in the scope of the protection.
Claims (9)
1. a kind of self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness, it is characterised in that comprise the following steps：
(1) antilemic plant system B and economical character excellent self-mating system A is hybridized, obtains hybridizing F1；
(2) the excellent F of economical character is chosen1Individual plant selfing, obtain selfing F2, epidemic disease band base of leaf bar identification in vitro is utilized in seedling stage
Method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC1F1；
(3) the excellent BC of economical character is chosen1F1Individual plant selfing, obtain selfing BC1F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC2F1；
(4) the excellent BC of economical character is chosen2F1Individual plant selfing, obtain selfing BC2F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC3F1；
(5) the excellent BC of economical character is chosen3F1Individual plant selfing, obtain selfing BC3F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC4F1；
(6) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC3F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC5F1；
(7) the excellent BC of economical character is chosen4F1Individual plant selfing, obtain selfing BC5F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then is returned with Elite inbred A, obtains first backcross generation BC6F1；
(8) the excellent BC of economical character is chosen6F1Individual plant selfing, obtain selfing BC6F2, in seedling stage using epidemic disease in vitro with base of leaf
Bar identification method carries out anti-epidemic disease identification, then individual plant selfing, obtains BC6F3, anti-epidemic disease is shown as from explanation, it is final to obtain height
Anti- epidemic disease and the excellent self-mating system of economical character；
(9) the high anti-epidemic disease obtained by above-mentioned steps and the excellent self-mating system of economical character are hybridized with other self-mating systems again,
High antilemic Dendrobidium huoshanness cenospecies can be selected.
2. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 1, it is characterised in that the agronomy
The excellent finger plant of character has one or more of states in high yield, high-quality, good stress resistance Breeding objective.
3. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 1, it is characterised in that the selection
Economical character excellent Dendrobidium huoshanness seedling when being selfed and being returned, the strain number of selection accounts for the 10-15% of total strain number.
4. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 3, it is characterised in that total strain
Number is no less than 50 plants.
5. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 1, it is characterised in that the epidemic disease
It is in vitro to be specially with base of leaf bar identification method：Same tether panel base of leaf bar is taken in seedling stage, 8-12min is rinsed with clear water, with 2-4% time
Sodium chlorate sterilizes 6-10min, finally rushes Xian 3-5 times with sterilized water, and be face-up put into sterilization is covered with double-deck moistening filter
In the culture dish of paper, it is (0.5-1.5) × 10 that surface, which sprays into concentration,5Individual/ml epidemic disease spore suspensions, be placed in after sealing 22 DEG C/
12h, 18 DEG C/12h incubator in alternation of light and darkness culture 2-4d, then carry out the identification of anti-epidemic disease and the statistical analysis of disease index.
6. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 5, it is characterised in that the anti-epidemic disease
The disease scale standard that disease is identified is：It is 0 grade, asymptomatic；1 grade, occur a small amount of epidemic disease shape scab on blade and scab is smudgy, disease
Spot area accounts for less than the 5% of the gross area；2 grades, occur obvious scab on blade, but it is scattered not in flakes, lesion area accounts for the gross area
6%-25%；3 grades, produce the epidemic disease shape scab of moderate quatity on blade and scab is linked to be piece, lesion area accounts for the gross area
26%-50%；4 grades, the epidemic disease Lesion number of blade is more, accounts for the 51%-75% of leaf area；5 grades, the epidemic disease scab on blade
Quantity is many and bisque is thicker, accounts for the 76%-100% of leaf area；
The statistical analysis technique of the disease index is：Disease index identification is calculated using the statistical analysis formula of disease index,
Standard of perfection is：Height is anti-, 0 ＜ disease index≤15；It is disease-resistant, 15 ＜ disease index≤35；Moderate resistance, 35 ＜ disease index≤55；
It is susceptible, 55 ＜ disease index≤75；Height sense, disease index >=75.
7. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 6, it is characterised in that the state of an illness
The statistical analysis formula of index is：DI=Σ (s × n) × 100/N × S, wherein, DI represents disease index, and s represents each state of an illness level
Other to represent numerical value, n represents each state of an illness rank disease number of sheets, and N represents investigation total number of sheets, and S represents highest level typical value.
8. the self-mating system breeding method of anti-epidemic disease evil Dendrobidium huoshanness according to claim 1, it is characterised in that the step
(9) other self-mating systems are specially the self-mating system with high anti-southern blight, the self-mating system with high polysaccharide or with the anti-anthrax of height in
The self-mating system of disease.
9. the self-mating system breeding method of Dendrobidium huoshanness is done harm to according to any described anti-epidemic diseases of claim 1-8, it is characterised in that choosing
Select whole tissue culture.
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| CN106358702A (en) * | 2016-08-30 | 2017-02-01 | 中国烟草总公司广东省公司 | Disease resistance intensity identifying method for resistance to tobacco anthracnose |
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| CN106358702A (en) * | 2016-08-30 | 2017-02-01 | 中国烟草总公司广东省公司 | Disease resistance intensity identifying method for resistance to tobacco anthracnose |
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