CN107182772A - Method and application of a kind of efficient seed selection without pollen contamination chrysanthemum - Google Patents
Method and application of a kind of efficient seed selection without pollen contamination chrysanthemum Download PDFInfo
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- CN107182772A CN107182772A CN201710428255.1A CN201710428255A CN107182772A CN 107182772 A CN107182772 A CN 107182772A CN 201710428255 A CN201710428255 A CN 201710428255A CN 107182772 A CN107182772 A CN 107182772A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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Abstract
The invention discloses a kind of method of efficient seed selection without pollen contamination chrysanthemum and application, this method includes：(1) according to chrysanthemum parent's pollen contamination degree and parent's reproductive development quality event, reasonable selection hybrid strain carries out artificial hybridization experiment；(2) sprouting quantity of the pollen on column cap in each combination is counted after artificial pollination；(3) the statistics full rate of ovary and setting percentage；(4) hybridization F is determined1For loose powder characteristic, analyze pollen contamination degree, so as to obtain obtaining no pollen contamination chrysanthemum filial generation, which part offspring can directly be promoted after overall merit as new varieties, and another part can be used as the intermediate materials without pollen contamination chrysanthemum breeding.Without Pollen plant highest rate it is 42% in the filial generation of this method seed selection the invention provides a kind of science, quick, efficient method of the seed selection without pollen contamination chrysanthemum, and the ratio of conventional method is typically not greater than 10%.Therefore, this method will provide theoretical direction to cultivate without pollen contamination chrysanthemum new varieties.
Description
Technical field
The invention belongs to chrysanthemum biology of reproduction and crossbreeding research field, it is related to a kind of efficient seed selection without pollen contamination
The method of chrysanthemum and application, relate in particular to seed selection after the discrepant Parent hybridization of pollen amount tool and obtain no pollen contamination chrysanthemum
Colored method and application.This method passes through with without pollen contamination (flower pesticide does not ftracture without pollen or anther wall) Dendranthema morifolium Varieties
' Kingfisher ', ' the beautiful tablet held before the breast by officials of southern agriculture ', ' Q1-62 ', ' QX-107 ', ' southern agriculture ice cream ' and ' QX-008 ' is female parent, ' Nan Nongfei
It is purple ', ' southern agriculture blood orange ', ' Nan Nongyin mountains ', ' southern agriculture prominence ' and ' Noah ' be that male parent is carried out 20 groups of artificial hybridization and tested；Statistics with
After analysis pollination 8 after sprouting quantity of the 24h pollen on column cap, pollination, 12 and 18 days full rates of ovary and setting percentages, synthesis comments
The hybridization fertility of each Parent of valency；By determining and analyzing hybridization F1For loose powder characteristic, including single inflorescence tubular flower number, single flower
Flower pesticide number, flower pesticide powder containing quantity, 5 characters of inflorescence powder containing quantity and apparent pollen amount, it is determined that each combination obtains no pollen contamination chrysanthemum
Flower F1For the ratio of strain.
Technical background
Chrysanthemum (Chrysanthemum morifolium) be perennial herb plant, the great tradition famous flower of China ten and
One of four gentleman in spending, possess long cultural and historical and abundant cultural deposits, meanwhile, because of its ornamental values and the economic values
Very high, demand is one of big Fresh Cutting flower in the world four greatly in cut-flower market at home and abroad.But chrysanthemum is in the process of blooming
In, the both sexes tubular flower in the middle of inflorescence is developed from foreign steamer to lubrication groove, and loose powder amount gradually increases, and substantial amounts of pollen load drops in ligule
Take, cause ornamental value to reduce, and excessive pollen is flown away in air, to the respiratory tract of human body especially to pollen hypersensitivity
Patient can produce pollinosis symptom, endanger human physical and mental health.Therefore, cultivating the chrysanthemum new varieties without pollen contamination has very
Important market application foreground,
For the seed selection of chrysanthemum new varieties, application at most maximally effective breeding method is sexual hybridization at present.It is used as seed
Plant, the compatibility of pollen and column cap, embryonic development and final setting percentage and the validity hybridized are direct in crossover process
Correlation, when carrying out a large amount of crossbreeding experiments, it should analyze the reproductive biology of selected parent.In addition,
During crossbreeding crossbreeding can be enormously simplify using without pollen contamination chrysanthemum without carrying out artificial emasculation for female parent
Program, it also avoid the injury that artificial emasculation is caused to inflorescence, be conducive to significantly improving breeding efficiency.Therefore, it is necessary to miscellaneous
The Parent fertility and F of friendship1Studied for loose powder characteristic, invent a kind of method of the efficient seed selection without pollen contamination chrysanthemum,
Excellent new varieties are provided for Market of Fresh Cut Flower, mitigate pollen contamination harm, while being also home decoration and other various fields
In conjunction in the reasonable disposition and Urbanization Construction of Fresh Cutting flower green plants configuration provides more more options.
The content of the invention
It is an object of the invention to provide a kind of method and its application of the efficient seed selection without pollen contamination chrysanthemum, it is desirably to obtain more
Many is applied in cut-flower market and afforestation without pollen contamination chrysanthemum new varieties, mitigates the harm of pollen contamination.
The purpose of the present invention can be achieved through the following technical solutions：
A kind of method of the efficient seed selection without pollen contamination chrysanthemum, comprises the following steps：
(1) using without pollen contamination Dendranthema morifolium Varieties as female parent, using pollen amount it is few, in, many Dendranthema morifolium Varieties as male parent enter pedestrian
Work hybridizes；
(2) sprouting quantity of the pollen on column cap in each cross combination is counted after artificial pollination；
(3) the statistics full rate of ovary and setting percentage；
(4) hybridization F is determined1For loose powder characteristic, including single inflorescence tubular flower number, single flower flower pesticide number, flower pesticide powder containing quantity, flower
5 characters of sequence powder containing quantity and apparent pollen amount, analyze pollen contamination degree, obtain without pollen contamination chrysanthemum filial generation.
Described in step (1) pollen or the indehiscent chrysanthemum product of anther wall are free of without pollen contamination Dendranthema morifolium Varieties for flower pesticide
Kind；Preferably flower pesticide is free of the Dendranthema morifolium Varieties of pollen；It is further preferred that Dendranthema morifolium Varieties of the flower pesticide without pollen are
' Kingfisher ', ' the beautiful tablet held before the breast by officials of southern agriculture ', ' Q1-62 '；The indehiscent Dendranthema morifolium Varieties of anther wall are ' QX-107 ', ' Nan Nongbing
Swash insult ', ' QX-008 '.
Male parent described in step (1) is preferably the few Dendranthema morifolium Varieties of pollen amount.The few Dendranthema morifolium Varieties of above-mentioned pollen amount are
' Noah '；Dendranthema morifolium Varieties in the pollen amount are ' southern agriculture prominence '；Dendranthema morifolium Varieties more than the pollen amount are ' Nan Nongfei is purple ',
' southern agriculture blood orange ', ' Nan Nongyin mountains '.
Sprouting quantity of the pollen on column cap in each cross combination is counted respectively in 24h after pollination in step (2)；Step
(3) in 8 after pollination, 12,18 days statistics ovary full number and not full number, calculate the full rate of ovary, treat seed maturity, unite
Count setting percentage；Ligulate flower number × 100% of the seed number of setting percentage (%)=average every inflorescence/average each inflorescence.
The method of artificial hybridization is described in step (1)：Cut off when maternal ligulate flower just reveals color ligulate flower petal until
The column cap of the inside is can see that, but column cap can not be brushed up against, and with sulfuric acid paper bag strictly bagging isolation；Simultaneously will be i.e. by loose powder
Male parent inflorescence carry out bagging isolation, the maternal column cap of observation is stretched out and during in " Y " font, and male parent is collected with writing brush in fine day
Fresh pollen, and pollen is lightly dipped on maternal ligulate flower pistil stigma, bagging isolation, after 1-2 days as stated above
Pollinate again.
3 inflorescences are collected in 24h after pollination in step (2), 5 little Hua are selected at random respectively, immediately with FAA (70% second
Alcohol:Glacial acetic acid:The volume ratio of formaldehyde is 90:5:5) it is fixed, it is stored in 4 DEG C of refrigerators standby；After 48h, 1molL is used-1NaOH
Solution softens 8h, and distilled water is cleaned 3 times, and is carefully stripped out style from little Hua, is placed on slide, is dripped with glue head
Pipe is added dropwise 0.1% aniline blue solution and dyed, and in fluorescence microscopy Microscopic observation and takes pictures immediately, records each pistil stigma
On pollen germination number, be repeated 15 times.
In step (3) 8 after artificial pollination, 12,18d take 5 inflorescences at random, visually observe the full situation of ovary, and
Full and not full number is counted, is taken pictures under stereomicroscope；Seed maturity is treated, inflorescence is cut, 60 flowers in each combining
Sequence is randomly divided into 3 parts, as 3 repetitions, each repeats 20 inflorescences, the ligulate flower numbers of 20 inflorescences is counted respectively and full
Seed number.
Described step (4) comprises the following steps：
(I) each F1The inflorescence for randomly select 3 well-growns for strain, being of moderate size, carries out tubular flower counting；
(II) randomly choose the tubular flower of 5 also non-loose powder respectively from above-mentioned 3 inflorescences, peeled off flower pesticide with dissecting needle
Out, each little Hua flower pesticide number is recorded；
(III) 60 pieces of flower pesticide are stripped from two three-wheel tubular flowers of the non-loose powder in periphery, are put into clean 10ml centrifuge tubes,
Dry in an oven, temperature is set to 50 DEG C, allow anther dehiscence to discharge pollen, after 3 days, smashing flower pesticide to pieces with dissecting needle uses up pollen
Amount sheds completely, then adds 6ml 20% (NaPO3)6Suspension, makes pollen scattered equal with WH-2 micro-whirlpool mixing vortexs
It is even, observed with microsyringe 2 μ L hanging drops of absorption on slide, under the microscope and carry out pollen amount counting, repeated
10 times, flower pesticide powder containing quantity=(pollen grain number × 3,000 on each slide)/60；
(IV) inflorescence powder containing quantity (grain/inflorescence)=single inflorescence tubular flower number × single flower flower pesticide number × flower pesticide powder containing quantity；
(V) in loose powder peak period, according to the volume size of pollen load, the collection complexity of pollen on column cap, apparent flower
Powder amount be divided into it is many, medium, less, without 4 grades, comparatively the substantial amounts of pollen load that is wherein scattered on column cap, collection are easy to simultaneously
And the very big kind divided into more than apparent pollen amount of collection capacity；Pollen load small volume, pollen collection are relatively easy on column cap
Divide the medium kind of apparent pollen amount into；And can only see a little pollen on column cap be classified as the few kind of apparent pollen amount；Have
Can not see pollen on a little kind column caps completely, it is open after hermaphrodite flower in flower pesticide keep complete for without apparent pollen amount kind.
Above-mentioned method is in seed selection without the application in pollen contamination chrysanthemum.
Method of the efficient seed selection without pollen contamination chrysanthemum provided by the present invention specifically includes following steps：
(1) with without pollen contamination (flower pesticide does not ftracture without pollen or anther wall) Dendranthema morifolium Varieties ' Kingfisher ', ' Nan Nong
Beautiful tablet held before the breast by officials ', ' Q1-62 ', ' QX-107 ', ' southern agriculture ice cream ' and ' QX-008 ' be female parent, ' Nan Nongfei is purple ', ' southern agriculture blood orange ', ' south
Agriculture silver mountain ', ' southern agriculture prominence ' and ' Noah ' be that male parent carries out 20 groups of artificial hybridization experiments, each kind flower pattern, posture all with compared with
High ornamental value.As shown in table 1,20 cross combinations are as shown in table 2 for hybrid strain pollen amount character.
The hybrid strain pollen amount character of table 1.
2. 20 cross combinations of table
(2) gather the little Hua of 3 inflorescences respectively in 24h after pollination, fluorescence microscopy Microscopic observation and record each combination
Sprouting quantity of the middle pollen on column cap, can filter out male parent and the preferable cross combination of maternal compatibility..
(3) in 8 after pollination, 12,18d respectively take 5 inflorescences, with the naked eye with stereomicroscope observation and record the full of ovary
Number and not full number.Seed maturity is treated, setting percentage is counted.The full rate of ovary has correlation with setting percentage, and setting percentage is with awarding
The ratio of 8 days full rates of ovary can intuitively show the relation of embryonic development and setting percentage after powder.
(4) hybridization F is determined1For loose powder characteristic, including single inflorescence tubular flower number, single flower flower pesticide number, flower pesticide powder containing quantity, flower
5 characters of sequence powder containing quantity and apparent pollen amount, analyze pollen contamination degree.Wherein flower pesticide powder containing quantity, inflorescence powder containing quantity and apparent
Pollen amount can be for judging the degree of pollen contamination, on this basis, and analysis obtains no pollen contamination F1It is more for strain
The character of parent, Rule Summary in combination.
Beneficial effects of the present invention：
The present invention for pollen amount character there is provided a kind of method of the efficient seed selection without pollen contamination chrysanthemum, with it is simple,
Fast and efficiently feature, is conducive to scientifically selecting hybrid strain, simplifies crossbreeding program, provides excellent for Market of Fresh Cut Flower
Good new varieties, while mitigating harm of the pollen contamination to human body.
Brief description of the drawings
Fig. 1 is combines 8 after 2 and the pollination of combination 11,12,18 days ovary forms.
Wherein, A, B, C be respectively combine 8 after 2 pollinations, 12 and 18 days full ovarys；D, E, F are respectively that combination 11 is pollinated
Afterwards 8,12 and 18 days not full ovarys.Scale：200μm.
Embodiment
1st, artificial hybridization
Ligulate flower petal is cut off when maternal ligulate flower just reveals color until can see that the column cap of the inside, but post can not be injured
Head, and with sulfuric acid paper bag strictly bagging isolation, fixed with safety pin, hang up label and date.Meanwhile, by i.e. by the father of loose powder
This inflorescence carries out bagging isolation.After about 2-3 days, the maternal column cap of observation stretches out and during in " Y " font, is received in fine day with writing brush
Collect the fresh pollen of male parent, and pollen is lightly dipped on maternal ligulate flower pistil stigma, bagging isolation, in label before
On write the maternal title of male parent and pollination time, pollinated again by method above after about 1-2 days.
2nd, sprouting of the pollen on column cap
3 inflorescences are collected in 24h after pollination, 5 little Hua are selected at random respectively, immediately with FAA (70% ethanol:Glacial acetic acid:
The volume ratio of formaldehyde is 90:5:5) it is fixed, it is stored in 4 DEG C of refrigerators standby.After 48h, 1molL is used-1NaOH solution softens 8h,
Distilled water is cleaned 3 times, and is carefully stripped out style from little Hua, is placed on slide, and 0.1% is added dropwise with glue head dropper
(m/v) aniline blue solution (0.1g aniline blues are added i.e. in 100ml deionized waters) is dyed, immediately in fluorescence microscope
Observe and take pictures under (Zeiss Axioskop 40), and record the pollen germination number on each pistil stigma, be repeated 15 times.
3rd, ovary form and setting percentage
8 after artificial pollination, take 5 inflorescences at random within 12,18 days, visually observe the full situation of ovary, and count full
With not full number, taken pictures under stereomicroscope.Seed maturity is treated, inflorescence is cut, 60 inflorescences divide at random in each combining
For 3 parts, as 3 repetitions each repeat 20 inflorescences, take back laboratory, and the ligulate flower number of 20 inflorescences is counted respectively and is satisfied
Full seed number.
4th, filial generation loose powder characteristic measurement
Single inflorescence tubular flower number：Each F1The inflorescence for randomly select 3 well-growns for strain, being of moderate size, is carried out
Tubular flower is counted.
Single flower flower pesticide number：Randomly choose the tubular flower of 5 also non-loose powder respectively from above-mentioned 3 inflorescences, will with dissecting needle
Flower pesticide is stripped out, and records each little Hua flower pesticide number.
Flower pesticide powder containing quantity：Strip 60 pieces of flower pesticide from two three-wheel tubular flowers of the non-loose powder in periphery, be put into clean 10ml from
In heart pipe, dry in an oven, temperature is set to 50 DEG C, allow anther dehiscence to discharge after pollen, about 3d, flower pesticide is smashed to pieces with dissecting needle
Pollen is shed completely as far as possible, then add 6ml 20% (m/v) (NaPO3)6Suspension (adds 20g in 100ml water
(NaPO3)6), pollen is uniformly dispersed with WH-2 micro-whirlpool mixing vortexs, drawing 2 μ L hanging drops with microsyringe is carrying
On slide, in the micro- Microscopic observations of OLYMPUS BX41 and pollen amount counting is carried out, be repeated 10 times, flower pesticide powder containing quantity=(each
Pollen grain number on slide × 3,000)/60.
Inflorescence powder containing quantity：Inflorescence powder containing quantity (grain/inflorescence)=single inflorescence tubular flower number × single flower flower pesticide number × flower pesticide contains
Powder amount.
Apparent pollen amount：In loose powder peak period, according to the volume size of pollen load, the collection complexity of pollen on column cap,
Apparent pollen amount be divided into it is many, medium, less, without 4 grades.Substantial amounts of pollen load, collection are wherein scattered on column cap comparatively
It is easy to and the very big kind divided into more than apparent pollen amount of collection capacity；Pollen load small volume, pollen collection phase on column cap
The kind medium to readily dividing apparent pollen amount into；And can only be seen on column cap a little pollen to be classified as apparent pollen amount few
Kind；Can not see pollen on some kind column caps completely, it is open after hermaphrodite flower in flower pesticide keep complete for without apparent pollen
Measure kind.
Interpretation of result
1st, sprouting of the pollen on column cap
In 20 cross combination, sprouting quantity of the pollen on column cap has larger difference.From table 3 it can be seen that
The pollen of combination 2, combination 3 and combination 19 sprouts quantity at most on column cap, respectively 19.93,17.53 and 15.63,
Most of pollen grain sticked on column cap has all been sprouted, and the pollen tube extending into style is high-visible, illustrates this
Compatibility between the parent of three combinations is good, available for later hybridization.The pollen of combination 8, combination 5 and combination 1 is in post
Quantity is sprouted on head medium, respectively 6.87,9.87 and 10.87 observe the pollen sticked on column cap in picture
Quantity is simultaneously few, but the pollen grain and many sprouted.And combine 10, combine 18 and the pollen of combination 11 sprouting number on column cap
Extremely low, respectively 0.96,0.45 and 0 are measured, only a small number of pollen tubes stretch into column cap on column cap, illustrate that these three are combined
Compatibility between parent is not high, there is crossing barrier.
Sprouting quantity of the 3 20 cross combination pollen of table on column cap
2nd, the full rate of ovary and setting percentage
After fertilization obstacle is one of principal element for causing Cross fertile rate low.From table 4 and accompanying drawing 1, combination 2, group
3, respectively 97.43%, 94.40% and 104.04% larger with 4 ratios of combination are closed, are illustrating these three combination embryonic developments just
Often, seldom occur Embryo Abortion, and combination 11, combination 15 and 16 ratios of combination are smaller, respectively 0,7.23% and 2.00% are said
Bright embryos most of in embryo development procedure there occurs abortion.
Discovery is further researched and analysed, to be hybridized without pollen Dendranthema morifolium Varieties as female parent, its Pistil recipient phase and knot
Real rate is relatively low；And have the kind of pollen grain using in not loose powder but flower pesticide as female parent, its Pistil recipient phase and setting percentage it is universal compared with
It is high.
4 20 full rates of cross combination ovary of table and setting percentage
3rd, F is hybridized1Seville orange flower powder pollution level is analyzed
Count 9 cross combination F1For FLORAL CHARACTERS OF BIG (table 5).319 strains are obtained in 9 cross combinations, have 57 to be
Offspring without pollen contamination, wherein do not have pollen in 23 strains in flower pesticide, 34 strains are that have pollen in flower pesticide but flower pesticide is not
Cracking causes that pollen can not be discharged.In addition, having counted each combination F1Ratio shared by Dai Zhongwu pollen contamination strains, combination 17
(' southern agriculture beautiful tablet held before the breast by officials ' × ' Noah ') obtains the ratio highest of no pollen contamination strain, accounts for 41.94%, combination 9 (' QX-008 ' ×
' Nan Nongyin mountains '), 19 (' Q1-62 ' × ' southern agriculture prominences ') of combination and combination 12 (' Kingfisher ' × ' Nan Nongfei purple ') offsprings
The middle ratio without pollen contamination strain is also higher, respectively 25.00%, 23.26% and 22.22%.
Further analysis is found, (to combine 12 as female parent without pollen kind, combine 17 and combination 19), in filial generation
High without pollen contamination strain proportion, mean height is up to 26.63%；And using polliniferous kind in not loose powder but flower pesticide as mother
This, it is low without pollen contamination strain proportion in filial generation, averagely only 8.70%.Therefore, using without pollen kind as female parent
Easily obtain no pollen contamination filial generation.In addition, using without pollen kind as female parent, it is many (combination 12) relative to pollen amount
Medium kind (combination 19) is as male parent with pollen amount, and the few kind of pollen amount is dirty without pollen in filial generation as male parent
Contaminate strain proportion higher, up to 41.94%.
The F of table 51Apparent pollen amount distribution of grades
Note：Underscore represents offspring's strain number without pollen in flower pesticide in bracket, and the right, which is represented, pollen but flower pesticide does not ftracture
Offspring's strain number.
Summarize
By combining miscellaneous with setting percentage and to 9 to sprouting of 20 cross combination pollen on column cap, the full rate of ovary
Hand over F1The analysis of seville orange flower powder pollution level, obtains a kind of method of the efficient seed selection without pollen contamination chrysanthemum, i.e., with without pollen chrysanthemum
Kind is hybridized as female parent using the kind that pollen amount is few as male parent, germination rate of the pollen on column cap and setting percentage compared with
It is low, but be easier to obtain without pollen contamination filial generation.This provides theoretical direction to cultivate without pollen contamination chrysanthemum new varieties,
Mitigate the harm of pollen contamination, while the also reasonable disposition for Fresh Cutting flower in home decoration and other various occasions and urbanization
The configuration provides of green plants more more options in construction.
Claims (10)
1. a kind of method of the efficient seed selection without pollen contamination chrysanthemum, it is characterised in that：Comprise the following steps：
(1) using without pollen contamination Dendranthema morifolium Varieties as female parent, using pollen amount it is few, in, many Dendranthema morifolium Varieties be male parent carry out manually it is miscellaneous
Hand over；
(2) sprouting quantity of the pollen on column cap in each cross combination is counted after artificial pollination；
(3) the statistics full rate of ovary and setting percentage；
(4) hybridization F is determined1Contain for loose powder characteristic, including single inflorescence tubular flower number, single flower flower pesticide number, flower pesticide powder containing quantity, inflorescence
5 characters of powder amount and apparent pollen amount, analyze pollen contamination degree, obtain without pollen contamination chrysanthemum filial generation.
2. according to the method described in claim 1, it is characterised in that：Described in step (1) without pollen contamination Dendranthema morifolium Varieties for flower
Medicine is free of pollen or the indehiscent Dendranthema morifolium Varieties of anther wall；Preferably flower pesticide is free of the Dendranthema morifolium Varieties of pollen；It is further preferred that
Dendranthema morifolium Varieties of the flower pesticide without pollen are ' Kingfisher ', ' the beautiful tablet held before the breast by officials of southern agriculture ', ' Q1-62 '；The anther wall does not ftracture
Dendranthema morifolium Varieties be ' QX-107 ', ' southern agriculture ice cream ', ' QX-008 '.
3. according to the method described in claim 1, it is characterised in that：Male parent described in step (1) is the few chrysanthemum of pollen amount
Kind.
4. the method according to claim 1 or 3, it is characterised in that：The few Dendranthema morifolium Varieties of the pollen amount are ' Noah '；Institute
It is ' southern agriculture prominence ' to state the Dendranthema morifolium Varieties in pollen amount；Dendranthema morifolium Varieties more than the pollen amount are ' Nan Nongfei is purple ', ' Nan Nonghong
Orange ', ' Nan Nongyin mountains '.
5. according to the method described in claim 1, it is characterised in that：Each hybridization group is counted respectively in 24h after pollination in step (2)
Sprouting quantity of the pollen on column cap in conjunction；In step (3) in 8 after pollination, 12,18 days statistics full numbers of ovary and not full
Number, calculates the full rate of ovary, treats seed maturity, counts setting percentage；The seed number of setting percentage (%)=average every inflorescence/average
Ligulate flower number × 100% of each inflorescence.
6. according to the method described in claim 1, it is characterised in that：The method of artificial hybridization is described in step (1)：In female parent
Ligulate flower cuts off ligulate flower petal until can see that the column cap of the inside, but can not brush up against column cap, and use sulfuric acid when just having revealed color
Paper bag strictly bagging isolation；The male parent inflorescence of loose powder will be subjected to bagging isolation simultaneously, the maternal column cap of observation stretches out simultaneously
And during in " Y " font, the fresh pollen of male parent is collected with writing brush in fine day, and pollen is lightly dipped in maternal ligulate flower gynoecium
On column cap, bagging isolation is pollinated again as stated above after 1-2 days.
7. method according to claim 1 or 5, it is characterised in that：3 inflorescences are collected in 24h after pollination in step (2),
Select 5 little Hua at random respectively, fixed immediately with FAA, be stored in 4 DEG C of refrigerators standby；After 48h, 1molL is used-1NaOH is molten
Liquid softens 8h, and distilled water is cleaned 3 times, and is carefully stripped out style from little Hua, is placed on slide, with glue head dropper
0.1% aniline blue solution is added dropwise to be dyed, in fluorescence microscopy Microscopic observation and takes pictures, is recorded on each pistil stigma immediately
Pollen germination number, be repeated 15 times.
8. method according to claim 1 or 5, it is characterised in that：In step (3) 8 after artificial pollination, 12,18d it is random
5 inflorescences are taken, the full situation of ovary is visually observed, and count full and not full number, are taken pictures under stereomicroscope；Wait to plant
It is sub ripe, inflorescence is cut, 60 inflorescences in each combination 3 parts are randomly divided into, as 3 repetitions each repeat 20 inflorescences,
The ligulate flower number and full seed number of 20 inflorescences are counted respectively.
9. according to the method described in claim 1, it is characterised in that：Described step (4) comprises the following steps：
(I) each F1The inflorescence for randomly select 3 well-growns for strain, being of moderate size, carries out tubular flower counting；
(II) randomly choose the tubular flower of 5 also non-loose powder respectively from above-mentioned 3 inflorescences, separated flower pesticide with dissecting needle
Come, record each little Hua flower pesticide number；
(III) 60 pieces of flower pesticide are stripped from two three-wheel tubular flowers of the non-loose powder in periphery, be put into clean 10ml centrifuge tubes, dried
Dried in case, temperature is set to 50 DEG C, allows anther dehiscence to discharge pollen, after 3 days, and smashing flower pesticide to pieces with dissecting needle makes pollen as far as possible complete
Shed entirely, then add 6ml 20% (NaPO3)6Suspension, makes pollen be uniformly dispersed with WH-2 micro-whirlpool mixing vortexs,
Observed with microsyringe 2 μ L hanging drops of absorption on slide, under the microscope and carry out pollen amount counting, repeat 10
It is secondary, flower pesticide powder containing quantity=(pollen grain number × 3,000 on each slide)/60；
(IV) inflorescence powder containing quantity (grain/inflorescence)=single inflorescence tubular flower number × single flower flower pesticide number × flower pesticide powder containing quantity；
(V) in loose powder peak period, according to the volume size of pollen load, the collection complexity of pollen on column cap, apparent pollen amount
Be divided into it is many, medium, less, without 4 grades, comparatively the substantial amounts of pollen load that is wherein scattered on column cap, collection are easy to and adopt
The very big kind divided into more than apparent pollen amount of collection amount；Pollen load small volume, pollen collection dividing into relatively easily on column cap
The medium kind of apparent pollen amount；And can only see a little pollen on column cap be classified as the few kind of apparent pollen amount；Some product
Can not see pollen on kind of column cap completely, it is open after hermaphrodite flower in flower pesticide keep complete for without apparent pollen amount kind.
10. the method in claim 1~9 described in any one is in seed selection without the application in pollen contamination chrysanthemum.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710428255.1A CN107182772A (en) | 2017-06-08 | 2017-06-08 | Method and application of a kind of efficient seed selection without pollen contamination chrysanthemum |
Applications Claiming Priority (1)
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| CN115707377A (en) * | 2022-11-30 | 2023-02-21 | 浙江海丰生物科技股份有限公司 | Method for optimizing breeding through chrysanthemum hybridization |
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| CN115707377A (en) * | 2022-11-30 | 2023-02-21 | 浙江海丰生物科技股份有限公司 | Method for optimizing breeding through chrysanthemum hybridization |
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