CN108271682A - A kind of STEVIA REBAUDIANA judicious mating producing method for seed - Google Patents
A kind of STEVIA REBAUDIANA judicious mating producing method for seed Download PDFInfo
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- CN108271682A CN108271682A CN201810047263.6A CN201810047263A CN108271682A CN 108271682 A CN108271682 A CN 108271682A CN 201810047263 A CN201810047263 A CN 201810047263A CN 108271682 A CN108271682 A CN 108271682A
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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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Abstract
The invention discloses a kind of STEVIA REBAUDIANA judicious mating production methods, belong to industrial crops production of hybrid seeds field.This method includes that the acquisition of the isolation implant of Parent plant, pollen preserves, preparation, liquid spray or the fluid drip of pollination liquid are pollinated and the collection of Stevia seed.According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, Cross fertile rate can be improved, ensure hybrid seed purity and improve seed yield rate.
Description
Technical field
The invention belongs to industrial crops production of hybrid seeds fields, and in particular to STEVIA REBAUDIANA judicious mating producing method for seed.
Background technology
STEVIA REBAUDIANA belongs to the not affine plant of selfing, and most kind self-pollinations are shaky, and seed is generated by hybridizing.
Existing STEVIA REBAUDIANA cross-breeding method is to arrive the cutting propagation seedling of each parent's strain or direct seeding seedling field planting
Same field is pollinated, phase mutual cross using media such as honeybees, to prepare STEVIA REBAUDIANA hybrid seed.These methods exist
Following shortcomings：Require there are flower synchronization that high, Cross fertile rate is low, seed purity is low, male parent between male parent and female parent
Plant waste is big, breeding cost is high.
In addition, different parents are colonized in same plot, because being natural crossing in existing STEVIA REBAUDIANA cross-breeding method
It hands over, Parent plant can mutually receive the pollen formation seed of other side by the pollination of the media such as honeybee.Due to sweet tea
The habits such as leaf chrysanthemum seed is light, has pappus, easily disperses everywhere with the wind after maturation, are difficult to exclude with economical character not when harvesting seed
Sufficient paternal plant is the incorporation that maternal reciprocal cross is formed by seed, leads to that seed purity is low, hybrid UV curing is high.
Simultaneously as the STEVIA REBAUDIANA same plant florescence continues one month or so more, the maturity period, difference was big between seed, Er Qietian
It easily disperses everywhere with the wind after leaf chrysanthemum seed maturity.Conventional Stevia seed collection method is when most of seed maturities one
Secondary property, which gathers in fall off seed after maternal plant is dried or manually shake plant in crop field separated in time, harvests seed.Former
Seed purity is higher obtained by seed harvesting method, be disadvantageous in that seed yield rate is low, maturity is inconsistent, germination percentage is low,
Later stage selected difficulty etc.；Kernel maturing consistency is preferable obtained by latter harvesting method, and seed production is high, but obtained seed is pure
Spend that low, hybrid UV curing is high, it is time-consuming, while easily plant is damaged and the later stage is caused to harvest in seed containing from injured branch
On the non-complete ripeness seed that falls off.Thus both harvesting approaches finally all cause to some extent hybrid seed purity it is low, germination
The deficiencies such as rate is low, production cost is high and later field plantation efficiency is low.
In addition, in other crop artificial pollination liquid announced at present prepare, required complicated components, such as patent
The pollination formula of liquid announced in 201610497946.2 includes just sucrose, glucose, boric acid, guar gum, soybean lecithin, 12
Ten kinds of compounds such as alkyl hydroxypropyl base phosphate, glycine betaine, potassium nitrate, ammonium chloride, choline chloride, increasing for component cause generally
Peasant household is difficult to enough effective popularizations purchased and accurately prepared, influence to pollinate success rate and technology.
Invention content
It is an object of the invention to：For the above problem present in STEVIA REBAUDIANA hybrid seeding, the present invention provides a kind of letter
Clean effective artificial pollination formula of liquid and a kind of STEVIA REBAUDIANA hybrid seed modern production technique.
The purpose of the present invention can be achieved through the following technical solutions：
One kind carrying out STEVIA REBAUDIANA judicious mating producing method for seed by liquid of pollinating, and this approach includes the following steps：
1) isolation implant of male parent and maternal plant；
2) acquisition of pollen preserves：In the male parent florescence, pollen, gained pollen for low temperature storage are acquired；
3) preparation of pollination liquid：Following substance is weighed by weight：D-araboascorbic acid sodium, Macrogol 4000, and
10~20 times of deionized waters of the two gross mass stir and evenly mix to obtain pollen buffer suspension liquid；Pollen is added in buffer suspension liquid,
It is configured to pollen buffer suspension liquid, obtains pollination liquid；
4) liquid spray or fluid drip pollination：After STEVIA REBAUDIANA maternal plant enters the florescence, daily or it is spaced 1-3 days,
The pollination liquid for taking step 3) is sprayed onto on new open maternal plant stylet head；
5) collection of Stevia seed：After pollination, using gauze bag by strain bagging or using liftoff 0~50cm in the ranks
It is laid with the modes such as gauze and collects mature seed.
In technical solution of the present invention：The isolation implant method of the pollen collection male parent and maternal plant includes but not
It is only limitted to zone isolation, is effectively isolated with modes such as gauze structure solariums, the isolation implant purpose of paternal plant is can
The acquisition of the consistent high-purity pollen of genotype is advantageously realized with the pollen collections such as bee, bumblebee biology in utilization, maternal plant
Isolation implant purpose is subsequent spray or instils when pollinating convenient for automation mechanized operation, saving pollen, raising pollination efficiency.
In technical solution of the present invention：The pollen collecting method includes artificial separation pollen and collects flower using honeybee
Powder.
In technical solution of the present invention：The acquisition period of little Hua used in the artificial separation pollen includes but are not limited to small
Flower with phyllary is contour, top is white will open when, should also include it is small the flowers are in blossom put front and back pollen with sprouting vigor times
Meaning period.
In technical solution of the present invention：The described period that pollen is acquired using honeybee is that small the flowers are in blossom puts rear pollen and have and sprout
The arbitrary period of vigor.
In technical solution of the present invention：The pollen collection time is morning 6：00~afternoon 6：00；
In technical solution of the present invention：It is various with sweetleaf chrysanthemum that the honeybee includes but are not limited to middle bee, bumblebee etc.
The bee class biology of powder acquisition capacity.
In technical solution of the present invention：Described further includes utilizing to include but are not limited to shedding plate using honeybee acquisition pollen
Equal tools are by the acquired pollen of honeybee when honeybee goes back to the nest under Automatic-falling.
In technical solution of the present invention：The pollen for low temperature storage temperature is -252 DEG C~4 DEG C.
In technical solution of the present invention：The mass ratio of the D-araboascorbic acid sodium and Macrogol 4000 be 0.01~
10：0.05~50.
In technical solution of the present invention：Pollen concentration is 20~100/microlitre in the pollen buffer suspension liquid.
In technical solution of the present invention：The fluid drip or liquid spray pollinated stage be when STEVIA REBAUDIANA maternal plant into
After entering the florescence (small the flowers are in blossom put), daily or interval progress in 1-3 days.
In technical solution of the present invention：The fluid drip or liquid spray pollination time is morning 6：00~afternoon 6：
00；
In technical solution of the present invention：The Stevia seed collection method is to utilize gauze bag by strain bagging or use
In the ranks liftoff 0~50cm is laid with the modes such as gauze and collects mature seed, and the aperture of gauze bag or gauze used is 0.1-2.0mm,
The harvesting method of seed may be used include but are not limited to dust catcher etc. machinery harvested to falling the seed being placed on gauze.
In technical solution of the present invention：The seed water content in harvest can be harvested once with separated in time, can also be waited
After all seeds are all ripe, using the method for vibration plant, all seeds are all shaken fall on gauze or in mesh bag after, disposably
Harvest.
Can be based on artificial, robot, mechanical arm and honeybee according to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention
Etc. auxiliary pollen oriented acquisition provide platform, improve pollen collection convenience, realize the oriented acquisition of target pollen, protect
Pollen purity is demonstrate,proved.
According to the present invention STEVIA REBAUDIANA judicious mating producing method for seed, can after the completion of pollen collection to paternal plant blade into
Harvest, the waste of paternal plant blade in effectively avoiding STEVIA REBAUDIANA hybrid seed from producing hybridize row to reduce STEVIA REBAUDIANA in time
Seed production cost.
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, the activity of excellent paternal pollen is ensured, has reduced excellent
The waste of matter pollen resource promotes application of the excellent pollen resource in crossbreeding；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, the production of STEVIA REBAUDIANA hybrid seed is made thoroughly to have broken away from by father
The control of maternal florescence consistency, realizes the independent assortment between Parent, for the production of more excellent combined hybrid seeds
Provide technical guarantee；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, alleviate in conventional method needed for pollen buffer
Complex component, improve pollination success rate and technology replicability；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, it is bad to reduce florescence high temperature, high humidity in conventional method
Influence of the environment to pollen activity improves the stability and controllability of the production of STEVIA REBAUDIANA hybrid seed；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, improves maternal plant little Hua orientations and receive paternal pollen
Probability, improve maternal plant little Hua Cross fertile rates；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, mixing from paternal plant seed is thoroughly eliminated,
It has ensured hybrid seed purity, has ensured the processing benefit of the planting benefit and subsequent extracted processing enterprise of peasant；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, the Stevia seed harvest used in conjunction with machinery for the later stage
Create condition, reduce early stage mature seed disperse everywhere caused by loss and STEVIA REBAUDIANA hybrid seed harvest in it is artificial
Expenditure, improves STEVIA REBAUDIANA hybrid seed productivity effect；
According to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, harvest can be reduced under the premise of ensureing seed harvest yield
Frequency reduces artificial expenditure, improves productivity effect；
It according to the STEVIA REBAUDIANA judicious mating producing method for seed of the present invention, once harvests, ensures after realizing all seed complete ripeness
The consistency of kernel maturing, improves percentage of seedgermination.
According to the present invention STEVIA REBAUDIANA judicious mating producing method for seed, emphasis solve in the STEVIA REBAUDIANA hybrid seed production of hybrid seeds with
Lower Railway Project：
1) it since present invention employs the isolation implants of male parent, collects and provides for high efficiency, the high-purity of pollen resource
Platform.
2) due to present invention employs the Cord blood of pollen, having ensured pollen activity, excellent pollen germplasm money is promoted
Effective application in source.
3) since present invention employs maternal isolation implants, for the automation in the spraying of later stage pollination liquid or drop hybridization
Operation provides condition, improves the crossing performance efficiency of pollen.
4) it due to the present invention provides easy pollen buffer method, improving the replicability of technology, ensures
Pollination success rate.
5) it since present invention employs the multiple bearing pollinations of artificial, robot or mechanical arm auxiliary, improves maternal small
Flower receives the probability of paternal pollen, improves Cross fertile rate.
6) due to present invention employs maternal isolation implant, having prevented with paternal plant to be mixing for maternal formation seed
Enter, the effective guarantee purity of hybrid seed.
7) since present invention employs maternal isolation implants, the receipts of STEVIA REBAUDIANA hybrid seed are carried out for later-stage utilization machinery
Obtain the condition of creating.
8) since present invention employs maternal isolation implants, being unified in liftoff 0~50cm in the ranks for the later stage is laid with gauze
Etc. modes collect mature seed and create condition.
9) due to present invention employs being laid with the modes such as gauze in the ranks to collect mature seed, being carried out for later-stage utilization machinery
The harvest of Stevia seed creates condition.
In conclusion as a result of STEVIA REBAUDIANA judicious mating production of hybrid seeds scheme, beneficial effects of the present invention include：
The present invention breaks through the limitation of traditional STEVIA REBAUDIANA hybrid seed production of hybrid seeds pattern, provides a kind of judicious mating production of hybrid seeds side
Method is to be selfed that shaky, kernel maturing is inconsistent including STEVIA REBAUDIANA, seed is light, tool pappus, it is ripe after easily with the wind everywhere
The production technology of the plant hybridization seed to disperse provides new direction；Paternal pollen utilization ratio can be improved, hybridization is solved
Seed production the limitations such as is influenced by Parent florescence consistency and hot and humid environment on pollen activity；It can ensure cenospecies
Sub- purity, maturity consistency and germination percentage etc.；It can solve artificial dependency degree height, seed matter in the production of STEVIA REBAUDIANA hybrid seed
Measure the problems such as unstable；Production cost can be reduced, production of hybrid seeds enterprise production of hybrid seeds benefit is improved；High quality seed can be produced, under guarantee
Swim the processing benefit of the planting benefit and extraction processing enterprise of peasant；Related scientific research achievement and agricultural machinery can preferably be pushed
Application in the production of STEVIA REBAUDIANA hybrid seed；STEVIA REBAUDIANA product can preferably be played in terms of improving human health
Effect, is conducive to the sustainable and healthy development of STEVIA REBAUDIANA industry.
It is mainly reflected in the following aspects using the modernization STEVIA REBAUDIANA hybrid seed production technology superiority of the present invention：
1) fixed for the pollen based on auxiliary such as artificial, robot, mechanical arm and honeybees by the isolation implant of paternal plant
Platform is provided to acquisition, improves pollen collection convenience, realizes the oriented acquisition of target pollen, ensure that pollen is pure
Degree；
It 2), can be to its leaf after paternal plant little Hua and its pollen collection by the isolation implant of paternal plant
Piece harvests in time, the waste of paternal plant blade in effectively avoiding STEVIA REBAUDIANA hybrid seed from producing, to reduce STEVIA REBAUDIANA hybridization
Seed production cost；
3) it by the prolonged cold preservation to pollen, has ensured the activity of excellent paternal pollen, has reduced high-quality pollen money
The waste in source promotes application of the excellent pollen resource in hybrid seeding；
4) by the prolonged cold preservation to pollen, the production of STEVIA REBAUDIANA hybrid seed is made thoroughly to break away from by the Parent florescence one
The limitation of cause property, realizes the independent assortment between Parent, and the production for more excellent combined hybrid seeds provides technology
It ensures；
5) by the prolonged cold preservation to pollen, reduce florescence high temperature in conventional method, high humidity poor environment to flower
The influence of powder vigor improves the stability and controllability of the production of STEVIA REBAUDIANA hybrid seed；
6) it provides and simplifies pollen buffer method, reduce the technical risk caused by multicomponent, improve technology
Stability and replicability；
7) the controllable orientation repeatedly pollination assisted by artificial, robot, mechanical arm, improves maternal plant little Hua orientations
The probability for receiving paternal pollen, improves Cross fertile rate；
8) it is planted by the physical isolation of maternal plant, thoroughly excludes being mixed into from paternal plant seed, effective guarantee
Hybrid seed purity improves later field planting benefit, ensures the processing of the planting benefit and subsequent extracted processing enterprise of peasant
Benefit；
9) mature seed is collected by being laid with the modes such as gauze in the ranks, the Stevia seed used in conjunction with machinery for the later stage
Harvest creates condition, and the artificial expenditure and early stage mature seed reduced in STEVIA REBAUDIANA hybrid seed harvest disperses led everywhere
The loss of cause improves STEVIA REBAUDIANA hybrid seed productivity effect；
10) due to by gauze bag by strain bagging or in the way of being in the ranks laid with gauze etc., can be in the premise for ensureing harvest yield
Lower reduction harvests frequency, reduces the artificial expenditure in STEVIA REBAUDIANA hybrid seed harvest, improves STEVIA REBAUDIANA hybrid seed productivity effect；
11) since by strain bagging or in the way of being in the ranks laid with gauze etc., it is all complete to realize all seeds by gauze bag
It is disposably harvested after maturation, has ensured kernel maturing consistency, improved percentage of seedgermination.
Specific implementation mode
With reference to embodiment, the present invention will be further described, and but the scope of the present invention is not limited thereto：
Embodiment 1
A kind of STEVIA REBAUDIANA judicious mating producing method for seed, includes the following steps：
Male parent material used is southern 1066 cutting propagation seedling of agriculture, and female parent material is your No. 1 cutting propagation seedling of sweet tea.
(1) acquisition of little Hua：After 1066 plant of southern agriculture enters florescence, in the morning 10：Before 00, picking will be opened and not opened
Little Hua liquid nitrogen is fixed -80 DEG C of postposition and is saved backup；
(2) preparation of pollen buffer suspension liquid：Weigh following substance, 10 parts of D-araboascorbic acid sodium, 50 parts of polyethylene glycol
4000 and the two gross mass 10 times of deionized waters, it is spare to obtain pollen buffer suspension liquid.
(3) preparation of pollination liquid：By weight 1 after taking step (1) preserved little Hua, freeze grinding to crush:50 are added flower
Powder buffer suspension liquid, is homogenized under condition of ice bath, through the strainer that aperture is 0.1mm after four layers of filtered through gauze, stands 30min, small
The heart discards supernatant, then pollen concentration is adjusted to 50 every microlitre with pollen buffer suspension liquid, obtains pollination liquid；
(4) artificial pollination：After sweet tea your No. 1 plant enters the florescence, the made pollen pollination liquid of step (3) is instilled into little Hua
On column cap, the time of instillation is selected in morning 6：00~morning 10：00 and afternoon 3：00~6：00, since initial bloom stage, often it bestowed by heaven
Powder is primary, until it is all it is small the flowers are in blossom discharge it is complete.
(5) bagging：Before artificial pollination and after pollination, the nylon net bag for being 0.8 millimeter with aperture carries out bagging isolation, seed
After maturation, setting percentage is investigated.
Comparative example 1-1
It is that its artificial pollination method is that pollen pollination liquid micro sprayer is small to the phase in full bloom with the difference of embodiment 1
Flower is pollinated, remaining operation is same as Example 1.
Comparative example 1-2
It is in pollen buffer suspension liquid only to contain D-araboascorbic acid sodium with the difference of embodiment 1.
Comparative example 1-3
It is to contain only Macrogol 4000 in pollen buffer suspension liquid with the difference of embodiment 1.
Comparative example 1-4
It is in pollen buffer suspension liquid containing 20 parts of D-araboascorbic acid sodium, 50 parts of polyethylene glycol with the difference of embodiment 1
4000
Comparative example 1-5
It is to contain in pollen buffer suspension liquid with the difference of embodiment 1：20 portions of sucrose, 10 parts of glucose, 4 parts of boric acid, 7 parts
Guar gum, 2 parts of soybean lecithins, 1.5 parts of dodecyl Hydroxypropyl phosphate ester glycine betaines, 1 part of potassium nitrate, 1 part of ammonium chloride, 0.5
Part choline chloride
Control group spontaneous pollination.
Following table is plant setting percentage obtained by each method, and as can be seen from the results, the STEVIA REBAUDIANA carried out according to the present invention is fixed
Cross fertile rate can be increased substantially to cross-breeding method.
Embodiment 2
A kind of STEVIA REBAUDIANA judicious mating producing method for seed, includes the following steps：
Male parent material used is southern 1066 cutting propagation seedling of agriculture, and female parent material is your No. 1 cutting propagation seedling of sweet tea.
(1) acquisition of little Hua：After 1066 plant of southern agriculture enters florescence, in the morning 10：Before 00, picking will be opened and not opened
Little Hua liquid nitrogen is fixed -120 DEG C of postposition and is saved backup；
(2) preparation of pollen buffer suspension liquid：Weigh following substance, 1 part of D-araboascorbic acid sodium, 50 parts of polyethylene glycol
4000 and the two gross mass 15 times of deionized waters, it is spare to obtain pollen buffer suspension liquid.
(3) preparation of pollination liquid：By weight 1 after taking step (1) preserved little Hua, freeze grinding to crush:50 are added flower
Powder buffer suspension liquid, is homogenized under condition of ice bath, through the strainer that aperture is 0.1mm after four layers of filtered through gauze, stands 30min, small
The heart discards supernatant, then pollen concentration is adjusted to 60 every microlitre with pollen buffer suspension liquid, obtains pollination liquid；
(4) artificial pollination：After sweet tea your No. 1 plant enters the florescence, the made pollen pollination liquid of step (3) is instilled into little Hua
On column cap, the time of instillation is selected in morning 6：00~morning 10：00 and afternoon 3：00~6：00, since initial bloom stage, often it bestowed by heaven
Powder is primary, until it is all it is small the flowers are in blossom discharge it is complete.
(5) bagging：Before artificial pollination and after pollination, the nylon net bag for being 0.8 millimeter with aperture carries out bagging isolation, seed
After maturation, setting percentage is investigated.
Comparative example 2-1
It is that its artificial pollination method is that pollen pollination liquid micro sprayer is small to the phase in full bloom with the difference of embodiment
Flower is pollinated, remaining operation is same as Example 2.
Comparative example 2-2
It is in pollen buffer suspension liquid only to contain D-araboascorbic acid sodium with the difference of embodiment 2.
Comparative example 2-3
It is to contain only Macrogol 4000 in pollen buffer suspension liquid with the difference of embodiment 2.
Comparative example 2-4
It is in pollen buffer suspension liquid containing 20 parts of D-araboascorbic acid sodium, 50 parts of polyethylene glycol with the difference of embodiment 2
4000
Comparative example 2-5
It is to contain in pollen buffer suspension liquid with the difference of embodiment 1：20 portions of sucrose, 10 parts of glucose, 4 parts of boric acid, 7 parts
Guar gum, 2 parts of soybean lecithins, 1.5 parts of dodecyl Hydroxypropyl phosphate ester glycine betaines, 1 part of potassium nitrate, 1 part of ammonium chloride, 0.5
Part choline chloride
Control group spontaneous pollination.
Following table is plant setting percentage obtained by each method, and as can be seen from the results, the STEVIA REBAUDIANA carried out according to the present invention is fixed
Cross fertile rate can be increased substantially to cross-breeding method.
Embodiment 3
A kind of STEVIA REBAUDIANA judicious mating producing method for seed, includes the following steps：
Male parent material used is southern 1066 cutting propagation seedling of agriculture, and female parent material is your No. 1 cutting propagation seedling of sweet tea.
(1) acquisition of little Hua：After 1066 plant of southern agriculture enters florescence, in the morning 10：Before 00, picking will be opened and not opened
Little Hua liquid nitrogen is fixed -200 DEG C of postposition and is saved backup；
(2) preparation of pollen buffer suspension liquid：Weigh following substance, 10 parts of D-araboascorbic acid sodium, 1 part of polyethylene glycol
4000 and the two gross mass 20 times of deionized waters, it is spare to obtain pollen buffer suspension liquid.
(3) preparation of pollination liquid：By weight 1 after taking step (1) preserved little Hua, freeze grinding to crush:50 are added flower
Powder buffer suspension liquid, is homogenized under condition of ice bath, through the strainer that aperture is 0.1mm after four layers of filtered through gauze, stands 30min, small
The heart discards supernatant, then pollen concentration is adjusted to 80 every microlitre with pollen buffer suspension liquid, obtains pollination liquid；
(4) artificial pollination：After sweet tea your No. 1 plant enters the florescence, the made pollen pollination liquid of step (3) is instilled into little Hua
On column cap, the time of instillation is selected in morning 6：00~morning 10：00 and afternoon 3：00~6：00, since initial bloom stage, often it bestowed by heaven
Powder is primary, until it is all it is small the flowers are in blossom discharge it is complete.
(5) bagging：Before artificial pollination and after pollination, the nylon net bag for being 0.8 millimeter with aperture carries out bagging isolation, seed
After maturation, setting percentage is investigated.
The difference of comparative example 3-1 and embodiment is that its artificial pollination method is by pollen pollination liquid ulv spraying
Device pollinates to phase little Hua in full bloom, remaining operation is same as Example 3.
Comparative example 3-2
It is in pollen buffer suspension liquid only to contain D-araboascorbic acid sodium with the difference of embodiment 2, remaining operation and reality
It is identical to apply example 2.
Comparative example 3-3
It is to contain only Macrogol 4000 in pollen buffer suspension liquid with the difference of embodiment 2, remaining operation and embodiment
It is identical.Comparative example 3-4
It is in pollen buffer suspension liquid containing 20 parts of D-araboascorbic acid sodium, 50 parts of polyethylene glycol with the difference of embodiment 3
4000, remaining operation is identical as embodiment.
Comparative example 3-5
It is to contain in pollen buffer suspension liquid with the difference of embodiment 1：20 portions of sucrose, 10 parts of glucose, 4 parts of boric acid, 7 parts
Guar gum, 2 parts of soybean lecithins, 1.5 parts of dodecyl Hydroxypropyl phosphate ester glycine betaines, 1 part of potassium nitrate, 1 part of ammonium chloride, 0.5
Part choline chloride, remaining operation are identical as embodiment.
Control group spontaneous pollination.
Following table is plant setting percentage obtained by each method, and as can be seen from the results, the STEVIA REBAUDIANA carried out according to the present invention is fixed
Cross fertile rate can be increased substantially to cross-breeding method.
Claims (4)
1. a kind of carrying out STEVIA REBAUDIANA judicious mating producing method for seed by liquid of pollinating, it is characterised in that：This approach includes the following steps：
1) isolation implant of male parent and maternal plant；
2) acquisition of pollen preserves：In the male parent florescence, pollen, gained pollen for low temperature storage are acquired；
3) preparation of pollination liquid：Following substance is weighed by weight：D-araboascorbic acid sodium, Macrogol 4000 and the two
10~20 times of deionized waters of gross mass stir and evenly mix to obtain pollen buffer suspension liquid；Pollen is added in buffer suspension liquid, prepares
At pollen buffer suspension liquid, pollination liquid is obtained；
4) liquid spray or fluid drip pollination：After STEVIA REBAUDIANA maternal plant enters the florescence, daily or it is spaced 1-3 days, takes step
Rapid pollination liquid 3) is sprayed onto on new open maternal plant stylet head；
5) collection of Stevia seed：After pollination, it is laid with by strain bagging or using liftoff 0~50cm in the ranks using gauze bag
The modes such as gauze collect mature seed.
2. STEVIA REBAUDIANA judicious mating producing method for seed according to claim 1, it is characterised in that：Cord blood temperature is -252
DEG C~4 DEG C.
3. STEVIA REBAUDIANA judicious mating producing method for seed according to claim 1, it is characterised in that：D-araboascorbic acid sodium and poly-
The mass ratio of ethylene glycol 4000 is 0.01~10：0.05~50.
4. STEVIA REBAUDIANA judicious mating producing method for seed according to claim 1, it is characterised in that：Pollen concentration is 20~100
Grain/microlitre pollen buffer suspension liquid.
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| CN201810047263.6A CN108271682B (en) | 2018-01-18 | 2018-01-18 | Directional hybridization seed production method for stevia rebaudiana |
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| CN102318549A (en) * | 2011-08-16 | 2012-01-18 | 南京农业大学 | Method with effects of saving cost and improving efficiency for liquid pollination of pear tree |
| CN103749288A (en) * | 2014-02-24 | 2014-04-30 | 何克勤 | Stevia rebaudiana mesh enclosure hybrid method |
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| KR20090132129A (en) * | 2008-06-20 | 2009-12-30 | 경북대학교 산학협력단 | New plant variety of stevia rebaudiana bertoni containing high rebaudioside-a and breeding method thereof |
| CN102318549A (en) * | 2011-08-16 | 2012-01-18 | 南京农业大学 | Method with effects of saving cost and improving efficiency for liquid pollination of pear tree |
| CN103749288A (en) * | 2014-02-24 | 2014-04-30 | 何克勤 | Stevia rebaudiana mesh enclosure hybrid method |
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