CN106106138B - A kind of red palm crossbreeding and rapid propagation method - Google Patents
A kind of red palm crossbreeding and rapid propagation method Download PDFInfo
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- CN106106138B CN106106138B CN201610683942.3A CN201610683942A CN106106138B CN 106106138 B CN106106138 B CN 106106138B CN 201610683942 A CN201610683942 A CN 201610683942A CN 106106138 B CN106106138 B CN 106106138B
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
A kind of red palm crossbreeding and rapid propagation method, successively including six selection, artificial pollination, aseptic seeding culture, Multiplying culture, strengthening seedling and rooting culture, test tube transplantation of seedlings steps.Red palm hybridization success rate can be improved using the inventive method, solve the problems, such as the advantages such as some cross combination flowering asynchronisms, quickly breeding high quality seedling, tool less investment, output height, to apply to large-scale production and the protection of red palm germ plasm resource.
Description
Technical field
The present invention relates to the breeding of plant and propagation method, refers in particular to a kind of red palm crossbreeding and quick breeding side
Method.
Background technology
The red palm（Anthurium adndraeanum）It is Araeceae Anthurium（Anthurium Schott ）Tropical grass
This flowers, its spend the peculiar gracefulness of appearance, spathe color it is beautiful it is different, be distributed widely in Central and South America.The red palm in the whole world there are about 19
Individual category or subgenus, possess initial species, mutation and cenospecies about more than 1000, be one of maximum category of Araeceae.It is red the palm due to
The unique spathe and florescence is extremely long, and there is high ornamental value, be cultivation in the world extensively, the basin of sales volume maximum cuts
One of flowers, at present the year amount of consumption of the red palm in the whole world be just introduced in me more than 10,000,000,000 RMB, the end of the eighties in last century
State simultaneously starts plant extensively；So far, China has formed single market the biggest in the world.
The red palm can generally use plant division and cutting propagation, and because red palm stem is short, tillering ability is also limited, therefore both nothings
The breeding coefficient of sexual reproduction method is not high, and reproduction speed is slow, although quite a few red palm seed can be by naturally miscellaneous
Hand over pollination and solid acquisition, but heterozygosis success rate is not high, and parental source is unknown, flowering asynchronism and due to each red palm variety seeds into
The ripe phase differs, and harvest time length, need to repeatedly harvest, repeatedly sow, less efficient, if red palm seed is sowed in natural environment or matrix
Sprouting, germination percentage are relatively low, and only 20%~30%.
The content of the invention
The present invention provides a kind of red palm crossbreeding and rapid propagation method, and its main purpose is to overcome the existing red palm miscellaneous
The defects such as conjunction rate is low, flowering asynchronism, breeding cycle is long, seed germination rate is low.
In order to solve the above technical problems, the present invention adopts the following technical scheme that：
A kind of red palm crossbreeding and rapid propagation method, comprise the following steps：
1) selection：Robust plant, the red palm for two kinds of different generas that resistance is excellent, ornamental value is high are chosen respectively as father
Originally it is and maternal；
2) artificial pollination：Through short-term low temperature（16 DEG C~18 DEG C）Stimulation or winter season, in flowering period, daystart collection father
This pollen, and artificial pollination is carried out to female parent, using the amber seed for being developed to maturation after pollination and not falling off as explant；
3) aseptic seeding culture：The outside rind of the amber seed is sloughed and cleaned up, first with concentration volume fraction
For alcohol-pickled 30~40 seconds of 74%~77%, then sterilized with the mercuric chloride solution that concentration mass fraction is 0.12%~0.18%
14~16 minutes, then with rinsed with sterile water 5~7 times, finally with filter paper blot the amber seed outer surface moisture and by its
It is inoculated on the germination medium that pH value is 5.5~6.0 and cultivates, is trained a height of 1~1.5 centimetre of plantlet；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.5~1.0g/L, inositol 80~
100mg/L, 1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.4mg/L, puridoxine hydrochloride (VB6) 0.4~
1.0mg/L, nicotinic acid 0.3~0.6mg/L, 6- benzyl aminoadenine（6-BA）0.1~0.5mg/L, methyl α-naphthyl acetate（NAA）0.005~
0.5 mg/L, 15~30g/L of sucrose, 4~5g/L of agar；
4) Multiplying culture：Step 3 will be passed through）In the plantlet that draws of culture cut root and be transferred to pH values for 5.5~
Cultivated in 6.0 proliferated culture medium, be trained the seedling that height is 1.5~3.0 centimetres；
The formula of proliferated culture medium is：MS minimal mediums, 80~100mg/L of inositol, 1.0~2.0mg/L of glycine,
Thiamine hydrochloride (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4~1.0mg/L, 6- benzyl aminoadenine（6-BA）
0.1~1.0 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, 4~5g/L of agar；
5) strengthening seedling and rooting culture：Step 4 will be passed through）In the seedling that draws of culture to be transferred to pH values strong for 5.5~6.0
Cultivated in seedling rooting culture medium, be trained a height of 3.0~5.0cm test tube seedling；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）0.5~3.0g/L, containing inositol 80
~100mg/L, 1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4
~1.0mg/L, methyl α-naphthyl acetate（NAA）0.005~0.5 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, agar 5~
6g/L；
6) test tube transplantation of seedlings：Step 5 will be passed through）In the test tube seedling that draws of culture be transferred in the greenhouse of shading more than 70%
Hardening 5~10 days, then it is taken out from strengthening seedling and rooting culture medium, the culture medium of root residual is cleaned, with 70% methyl Tobe
Plant in seedling medium and cultivated into seedling after the 800 times of liquid disinfectant processing in Tianjin or carbendazol wettable powder；
The formula of seedling medium is：It is 8 by volume by peat soil, perlite, river sand:1:1~9:0.5:0.5 mixing shape
Into mixed-matrix.
Further, step 2）Middle artificial pollination process, when male parent is identical with the maternal florescence, maternal spadix gynoecium
Start secreting mucus 1~2 day；Paternal pollen is taken directly to be invested with writing brush in maternal column cap in daystart 4 points~6.
Further, step 2）In artificial pollination, when male parent and maternal flowering asynchronism, male parent material is taken to be placed in artificial gas
Wait in room, wherein night sets 16 DEG C~18 DEG C of temperature, daytime 28 DEG C~32 DEG C of growing temperatures, illuminance 2000lx, during illumination
Long 12h/ d；Processing more than two weeks untill paternal pollen is collected, directly invests pollen on maternal post with writing brush afterwards
On head.
Further, step 3）~5）In cultivation temperature be 22~27 DEG C.
Further, step 3）~5）In intensity of illumination be 1500~2000lx.
Further, step 3）~5）Light irradiation time is 10~14 h/d.
Further, step 6）In intensity of illumination be 1500~2500lx.
Further, the kind of the red palm can be Anthurium andraeanum（Great Ye fancy candles lit in the bridal chamber at wedding）、A.bakeri
（Green spathe fancy candles lit in the bridal chamber at wedding）, A.crystallinum, A.dussii (triangle leaf fancy candles lit in the bridal chamber at wedding), A.holtonianum（Anistree leaf flower
Candle）、A.hookeri、A.kempteri、A.longipetiolatum、A.macrolobum、A.magnificure、
A.miquelianum、A.nymphillium、A.ozfisianum、A.pedatoradiatum、A.putamayo、
A.scanden、A.signature（Trilobated leaf fancy candles lit in the bridal chamber at wedding）、A.subsignatum、A.undatum、A.variabile、
A.veitchii、A.warocquenum。
In the present invention, germination medium, proliferated culture medium, the MS that uses in strengthening seedling and rooting culture medium cultivate substantially or
1/2MS minimal mediums provide the inorganic nutritive elements such as N, P, K for the growth of red palm seedling, to meet the base of its growth
This demand.
The activated carbon used in the present invention, English name activated carbon (abbreviation AC), have and adsorb the red palm
The effect for the noxious material being metabolized in growth.
The inositol used in the present invention, English name Inositol, it works in the mutual conversion of carbohydrate, is thin
The structure material of cell wall.Inositol participates in carbohydrate, the physiological activity such as phospholipid metabolism and ionic equilibrium, has and helps active matter
The effect that matter plays a role, callus growth can be promoted, the formation to embryoid and bud also has good facilitation.
The glycine used in the present invention, English name Glycine, it is possible to increase stress resistance of plant, it is special to plant growth
It is not that photosynthesis has unique facilitation；It can increase the chlorophyll content of plant, improve the activity of enzyme, promote two
The infiltration of carbonoxide, make photosynthesis more vigorous, so as to improve the quality of the red palm.
The thiamine hydrochloride (abbreviation VB1) used in the present invention, is a kind of existence form of vitamin B1, thiamine hydrochloride
Induction for anthurium andraeanum callus has facilitation, moreover it is possible to promotes cell division, improves its speed of growth.
The puridoxine hydrochloride (abbreviation VB6) used in the present invention, alias vitamin B6, can promote plant root growth,
Improve the speed of growth of the red palm.
The nicotinic acid used in the present invention is also referred to as vitamin B3, or nicotinic acid, molecular formula：C6H5NO2.Its major physiological
Effect is to participate in carbohydrate metabolism, phospholipid metabolism and ionic equilibrium effect have facilitation to tissue fast-growth, right
The formation of embryoid and bud has good influence.
The 6- benzyl aminoadenines used in the present invention, English name 6-Benzylaminopurine (abbreviation 6-BA),
Belong to broad spectrum activity plant growth regulator, red palm cell growth can be promoted, suppress the degraded of its chlorophyll, improve containing for amino acid
Amount, Delaying Leaf-Senescence etc..
The methyl α-naphthyl acetate used in the present invention, English name 1-Naphthaleneacetic acid (abbreviation NAA), it is wide
Spectral pattern plant growth regulator, cell division can be promoted with expanding, induced synthesis adventitious root increase fruit setting, prevent shedding, change
Female, male flower ratio etc..
The sucrose used in the present invention plays a part of energy substance and Osmolyte regulator, and in addition to energy supply, moreover it is possible to
Evoked callus tissue breaks up again.
Compared to the prior art, beneficial effect caused by the present invention is:
1st, the red palm produced by the invention has hybridization success rate high, emergence fast, robust seedling, seedling quality better, well-grown etc.
Feature, can solve the problems, such as some cross combination flowering asynchronisms, quickly breeding high quality seedling, and be produced on a large scale have and throw
Enter less, the advantage such as output height.A large amount of seedlings can be obtained, for large-scale production and the protection of red palm germ plasm resource.
2nd, in various composition of the invention, minimal medium only ensures the existence of culture and minimum physiological activity, flesh
Alcohol, thiamine hydrochloride promote callus growth, activated carbon, glycine, puridoxine hydrochloride, nicotinic acid, 6- benzyls aminoadenine, naphthalene
Acetic acid can promote cell division, make tissue rapid growth so that the red palm can quickly be bred.
3rd, at present, though having red palm aseptic seeding and test tube seedling breeding method both at home and abroad, can be adapted to without a kind of culture medium
In the aseptic seeding and test tube seedling of the red palm of different cultivars, therefore the present invention has larger advantage in adaptability, can be applicable
In the red palm of each kind.
Embodiment
Illustrate the embodiment of the present invention below.
A kind of red palm crossbreeding and rapid propagation method, comprise the following steps：
Step 1：Selection
Robust plant, the red palm for two kinds of different generas that resistance is excellent, ornamental value is high are chosen respectively as male parent and mother
This.The kind of the red palm can be Anthurium andraeanum（Great Ye fancy candles lit in the bridal chamber at wedding）、A.bakeri（Green spathe fancy candles lit in the bridal chamber at wedding）、
A.crystallinum, A.dussii (triangle leaf fancy candles lit in the bridal chamber at wedding), A.holtonianum（Anistree leaf fancy candles lit in the bridal chamber at wedding）、A.hookeri、
A.kempteri、A.longipetiolatum、A.macrolobum、A.magnificure、A.miquelianum、
A.nymphillium、A.ozfisianum、A.pedatoradiatum、A.putamayo、A.scanden、A.signature
（Trilobated leaf fancy candles lit in the bridal chamber at wedding）、A.subsignatum、A.undatum、A.variabile、A.veitchii、A.warocquenum.
Step 2：Artificial pollination
Through short-term low temperature（16 DEG C~18 DEG C）Stimulation or winter season, in flowering period, daystart gathers the pollen of male parent, and
Artificial pollination is carried out to female parent, using the amber seed for being developed to maturation after pollination and not falling off as explant；
During artificial pollination, when male parent is identical with the maternal florescence, maternal spadix gynoecium start secreting mucus 1~
2 days；Paternal pollen is taken directly to be invested with writing brush in maternal column cap in daystart 4 points~6；
When male parent and maternal flowering asynchronism, male parent material is taken to be placed in phjytotron, wherein night sets temperature 16
DEG C~18 DEG C, daytime 28 DEG C~32 DEG C of growing temperatures, illuminance 2000lx, light irradiation time 12h/ d；Processing is more than two weeks, directly
Untill paternal pollen is collected, pollen is directly invested in maternal column cap with writing brush afterwards.
Step 3：Aseptic seeding culture
The outside rind of the amber seed is sloughed and cleaned up, first with the wine that concentration volume fraction is 74%~77%
Essence immersion 30~40 seconds, then with concentration mass fraction be 0.12%~0.18% mercuric chloride solution sterilize 14~16 minutes, then
With rinsed with sterile water 5~7 times, finally blot the moisture of the amber seed outer surface with filter paper and be inoculated into pH value as 5.5
Cultivated on~6.0 germination medium, be trained a height of 1~1.5 centimetre of plantlet；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.5~1.0g/L, inositol 80~
100mg/L, 1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.4mg/L, puridoxine hydrochloride (VB6) 0.4~
1.0mg/L, nicotinic acid 0.3~0.6mg/L, 6- benzyl aminoadenine（6-BA）0.1~0.5mg/L, methyl α-naphthyl acetate（NAA）0.005~
0.5 mg/L, 15~30g/L of sucrose, 4~5g/L of agar；
The effect of germination medium is to improve the germination percentage and speed of germination of seed, and it is small plant to make seed Fast Growth
Strain, reduces the waste of resource, amber seed is cultivated in germination medium can be trained a height of 1~1.5 centimetre for 30~35 days
Plantlet.
Step 4：Multiplying culture
The plantlet drawn by the culture in step 3 is cut into root and is transferred to the Multiplying culture that pH values are 5.5~6.0
Cultivated in base, be trained the seedling that height is 1.5~3.0 centimetres；
The formula of proliferated culture medium is：MS minimal mediums, 80~100mg/L of inositol, 1.0~2.0mg/L of glycine,
Thiamine hydrochloride (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4~1.0mg/L, 6- benzyl aminoadenine（6-BA）
0.1~1.0 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, 4~5g/L of agar；
The effect of proliferated culture medium is that the efficiency of production is improved to mass produce, and plantlet is in proliferated culture medium
Culture 30~60 days, you can be trained the seedling that height is 1.5~3.0 centimetres.
Step 5：Strengthening seedling and rooting culture
The seedling drawn by the culture in step 4 is transferred to pH values to be trained in 5.5~6.0 strengthening seedling and rooting culture mediums
Support, be trained a height of 3.0~5.0cm test tube seedling；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）0.5~3.0g/L, containing inositol 80
~100mg/L, 1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4
~1.0mg/L, methyl α-naphthyl acetate（NAA）0.005~0.5 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, agar 5~
6g/L；
The effect of strengthening seedling and rooting culture medium is to improve the survival rate of test tube seedling, and promotes its growth.Seedling is in strengthening seedling and rooting
Cultivated 30~40 days in culture medium, you can be trained a height of 3.0~5.0cm test tube seedling.
Step 6：Test tube transplantation of seedlings
The test tube seedling drawn by the culture in step 5 is transferred to hardening 5~10 days in the greenhouse of shading more than 70%, so
It is taken out from strengthening seedling and rooting culture medium afterwards, cleans the culture medium of root residual, it is wettable with 70% thiophanate methyl or carbendazim
Property pulvis 800 times of liquid disinfectants processing after plant in seedling medium cultivation into seedling；
The formula of seedling medium is：It is 8 by volume by peat soil, perlite, river sand:1:1~9:0.5:0.5 mixing shape
Into mixed-matrix；
Cultivated 45~90 days in seedling medium, you can cultivate into seedling.
Wherein, the cultivation temperature in step 3~five is 22~27 DEG C, and intensity of illumination is 1500~2000lx, light irradiation time
For 10~14 h/d.Intensity of illumination in step 6 is 1500~2500lx.Red palm seedling produced by the invention, which has, to be hybridized to
The features such as power is high, emergence is fast, robust seedling, seedling quality better, well-grown, and be produced on a large scale, there is less investment, output
High advantage.A large amount of seedlings can be obtained, protection and merchandized handling for red palm germ plasm resource.At present, though having both at home and abroad
Red palm aseptic seeding and test tube seedling breeding method, but without a kind of culture medium be suitably adapted for the aseptic seeding of the red palm of different cultivars with
Test tube seedling, therefore the present invention has larger advantage in adaptability, can be suitably used for the red palm of each kind.
In addition, in the various composition of the present invention, minimal medium only ensures the existence of culture and minimum physiological activity,
Inositol, thiamine hydrochloride callus growth, activated carbon, glycine, puridoxine hydrochloride, nicotinic acid, 6- benzyls aminoadenine, naphthalene second
Acid can promote cell division, make tissue rapid growth so that the red palm can quickly be bred.
Embodiment one
Step 1：Selection
Choose mobile telephone（Dakota）With it is bright cherry-red（Cherry red）, and it is greatly maternal with eldest brother, bright cherry-red is male parent.
Step 2：Artificial pollination
In December~2 month flowering period, pollen is gathered when bright cherry-red spadix pollen bag ftractures 1~2 day, uses writing brush
In the spadix base portion secretion mucus for being invested maternal mobile telephone.Bagging after pollination, hangs up label, indicates Parent, miscellaneous
The information such as date are handed over, wait collection during seed maturity to yellowing to be used as explant；
The success rate of pollination is counted after 4 months, up to more than 90%.
Step 3：Aseptic seeding culture
With washing powder by amber seed surface contaminants rinsed clean, outside rind is pumped and cleaned up afterwards, used
75% alcohol-pickled 35 seconds, then sterilized 15 minutes with 0.15% mercuric chloride solution, rinsed with sterile water 6 times, will after filter paper suck dry moisture
Amber seed is inoculated on germination medium, and seed, which starts to sprout, at the 13rd day shows money or valuables one carries unintentionally, and seed cotyledons germinate at the 25th day, and the 35th
It can form 1~1.5 centimetre of high plantlet；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.5g/L, inositol 100mg/L, sweet ammonia
Sour 2.0mg/L, thiamine hydrochloride (VB1) 0.4mg/L, the mg/L of puridoxine hydrochloride (VB6) 0.5, nicotinic acid 0.5 mg/L, 6- benzyl ammonia
Base adenine（6-BA）0.5mg/L, methyl α-naphthyl acetate（NAA）0.05 mg/L, sucrose 20g/L, the g/L of agar 5, its pH value are 6.0.
Step 4：Multiplying culture
It is transferred to after plantlet is cut into root on proliferated culture medium, it is small for 1.5~3.0 centimetres that height can be formed after 60 days
Seedling；
The formula of proliferated culture medium is：MS minimal mediums, inositol 100mg/L, glycine 2.0mg/L, thiamine hydrochloride
(VB1) 0.4mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, 6- benzyl aminoadenine（6-BA）0.2 mg/L, the mg/ of nicotinic acid 0.5
L, sucrose 30g/L, agar 5g/L, its pH value are 6.0.
Step 5：Strengthening seedling and rooting culture
Seedling is transferred in strengthening seedling and rooting culture medium and cultivated, the high test tube seedlings of 3.0~5.0cm can be formed after 30 days；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）1.5g/L, inositol 100mg/L,
Glycine 2.0mg/L, thiamine hydrochloride (VB1) 0.4mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, nicotinic acid 0.5mg/L, naphthalene second
Acid（NAA）0.05mg/L, sucrose 25g/L, agar 5g/L, its pH value are 6.0.
Step 6：Test tube transplantation of seedlings
Test tube seedling is placed in hardening 7 days in the greenhouse of shading more than 70%, then cleans root remaining medium, then with 70%
Planted after the 800 times of liquid disinfectants processing of thiophanate methyl or carbendazol wettable powder into peat soil, perlite, river sand（Volume ratio
8:1:1）Mixed-matrix in cultivate 60 days, obtain seedling.
Condition of culture is 24 DEG C, illuminance 2000lx, light irradiation time 12h/ d of cultivation temperature in above-mentioned steps three~five；On
It is intensity of illumination 2000lx to state condition of culture in step 6.Most afterwards through statistics, survival rate 98%.
Embodiment two
Step 1：Selection
Choose Pola（Baby pear）And illusion（Princess Amalia Brilliant）, and using Pola as female parent,
Illusion is male parent.
Step 2：Artificial pollination
Because Pola and dreamlike florescence differ, male parent illusion parent material is taken to be placed in phjytotron, wherein night
Set 16 DEG C of temperature, daytime 28 DEG C~32 DEG C of growing temperatures, illuminance 2000lx, light irradiation time 12h/ d；Processing 1 month；
Dreamlike spadix pollen bag gathers pollen when ftractureing 1~2 day, and the spadix base section of maternal Pola is invested with writing brush
Secrete in mucus.Bagging after pollination, hangs up label, the information such as Parent, hybridization date is indicated, when waiting seed maturity to yellowing
Collection is used as explant；
The success rate of pollination is counted after 4 months, up to more than 75%.
Step 3：Aseptic seeding culture
With washing powder by amber seed surface contaminants rinsed clean, outside rind is pumped and cleaned up afterwards, used
74% alcohol-pickled 30 seconds, then sterilized 14 minutes with 0.12% mercuric chloride solution, rinsed with sterile water 5 times, will after filter paper suck dry moisture
Amber seed is inoculated on germination medium, and seed, which starts to sprout, at the 10th day shows money or valuables one carries unintentionally, and seed cotyledons germinate at the 20th day, and the 30th
It can form 1~1.5 centimetre of high plantlet；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.7g/L, inositol 80mg/L, sweet ammonia
Sour 2.0mg/L, thiamine hydrochloride (VB1) 0.5mg/L, the mg/L of puridoxine hydrochloride (VB6) 0.5, nicotinic acid 0.5 mg/L, 6- benzyl ammonia
Base adenine（6-BA）0.5 mg/L, methyl α-naphthyl acetate（NAA）0.05 mg/L, sucrose 20g/L, the g/L of agar 5, its pH value are 5.6.
Step 4：Multiplying culture
It is transferred to after plantlet is cut into root on proliferated culture medium, it is small for 1.5~3.0 centimetres that height can be formed after 40 days
Seedling；
The formula of proliferated culture medium is：MS minimal mediums, inositol 80mg/L, glycine 2.0mg/L, thiamine hydrochloride
(VB1) 0.5mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, 6- benzyl aminoadenine（6-BA）0.5mg/L, the mg/L of nicotinic acid 0.5,
Sucrose 30g/L, agar 5g/L, its pH value are 5.6.
Step 5：Strengthening seedling and rooting culture
Seedling is transferred in strengthening seedling and rooting culture medium and cultivated, the high test tube seedlings of 3.0~5.0cm can be formed after 30 days；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）1.0g/L, inositol 100mg/L,
Glycine 2.0mg/L, thiamine hydrochloride (VB1) 0.4mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, nicotinic acid 0.5mg/L, naphthalene second
Acid（NAA）0.05mg/L, sucrose 25g/L, agar 5g/L, its pH value are 5.6.
Step 6：Test tube transplantation of seedlings
Test tube seedling is placed in hardening 5 days in the greenhouse of shading more than 70%, then cleans root remaining medium, then with 70%
Planted after the 800 times of liquid disinfectants processing of thiophanate methyl or carbendazol wettable powder into peat soil, perlite, river sand（Volume ratio
8:1:1）Mixed-matrix in cultivate 70 days, obtain seedling.
Condition of culture is 24 DEG C, illuminance 2000lx, light irradiation time 13h/ d of cultivation temperature in above-mentioned steps three~five；On
It is intensity of illumination 2200lx to state condition of culture in step 6.Most afterwards through statistics, survival rate 95%.
Embodiment three
Step 1：Selection
Choose Pandora（Bandola）And rainbow conk（YunZhi）, and using Pandora as female parent, rainbow conk is male parent.
Step 2：Pollination
Because Pandora and rainbow conk florescence differ, rainbow conk parent material is taken to be placed in phjytotron, wherein night sets
Put 18 DEG C of temperature, daytime 28 DEG C~32 DEG C of growing temperatures, illuminance 2000lx, light irradiation time 12h/ d；Processing 45 days；In rainbow conk
Red spadix pollen bag gathers pollen when ftractureing 1~2 day, and the spadix base portion that maternal mobile telephone is invested with writing brush sticks
In liquid.Bagging after pollination, hangs up label, indicates the information such as Parent, hybridization date, is gathered when waiting seed maturity to yellowing
As explant；
The success rate of pollination is counted after 4 months, up to more than 70%.
Step 3：Aseptic seeding culture
With washing powder by amber seed surface contaminants rinsed clean, outside rind is pumped and cleaned up afterwards, used
77% alcohol-pickled 40 seconds, then sterilized 16 minutes with 0.18% mercuric chloride solution, rinsed with sterile water 7 times, will after filter paper suck dry moisture
Amber seed is inoculated on germination medium, and seed, which starts to sprout, at the 15th day shows money or valuables one carries unintentionally, and seed cotyledons germinate at the 25th day, and the 30th
It can form 1~1.5 centimetre of high plantlet；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.5g/L, inositol 90mg/L, sweet ammonia
Sour 2.0mg/L, thiamine hydrochloride (VB1) 0.4mg/L, the mg/L of puridoxine hydrochloride (VB6) 0.4, nicotinic acid 0.5 mg/L, 6- benzyl ammonia
Base adenine（6-BA）0.5 mg/L, sucrose 20g/L, agar 5g/L, its pH value are 5.8.
Step 4：Multiplying culture
It is transferred to after plantlet is cut into root on proliferated culture medium, it is small for 1.5~3.0 centimetres that height can be formed after 45 days
Seedling；
The formula of proliferated culture medium is：MS minimal mediums, inositol 80mg/L, glycine 2.0mg/L, thiamine hydrochloride
(VB1) 0.5mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, 6- benzyl aminoadenine（6-BA）0.8mg/L, methyl α-naphthyl acetate（NAA）
0.02mg/L, nicotinic acid 0.5 mg/L, sucrose 30g/L, agar 5g/L, its pH value are 5.8.
Step 5：Strengthening seedling and rooting culture
Seedling is transferred in strengthening seedling and rooting culture medium and cultivated, the high test tube seedlings of 3.0~5.0cm can be formed after 40 days；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）1.5g/L, inositol 100mg/L,
Glycine 2.0mg/L, thiamine hydrochloride (VB1) 0.4mg/L, puridoxine hydrochloride (VB6) 0.5mg/L, nicotinic acid 0.5mg/L, naphthalene second
Acid（NAA）0.05 mg/L, sucrose 30g/L, agar 5g/L, pH 5.8.
Step 6：Test tube transplantation of seedlings
Test tube seedling is placed in hardening 5 days in the greenhouse of shading more than 70%, then cleans root remaining medium, then with 70%
Planted after the 800 times of liquid disinfectants processing of thiophanate methyl or carbendazol wettable powder into peat soil, perlite, river sand（Volume ratio
8:1:1）Mixed-matrix in cultivate 80 days, obtain seedling.
Condition of culture is 24 DEG C, illuminance 2000lx, light irradiation time 10h/ d of cultivation temperature in above-mentioned steps three~five；On
It is intensity of illumination 2500lx to state condition of culture in step 6.Most afterwards through statistics, survival rate 92%.
With being trained 7 using soil plantation the time required to drawing the seedling that 7~9cm is trained using the present invention through overtesting
Contrast table the time required to~9cm seedling is as follows：
It can thus be appreciated that the seedling required time that 7~9cm is trained using the present invention is trained 7 compared to using soil plantation
To be greatly shortened the time required to~9cm seedling.
The embodiment of the present invention is above are only, but the design concept of the present invention is not limited thereto, it is all to utilize this
Conceive the change that unsubstantiality is carried out to the present invention, the behavior for invading the scope of the present invention all should be belonged to.
Claims (6)
1. a kind of red palm crossbreeding and rapid propagation method, it is characterised in that comprise the following steps：
1) selection：Robust plant, the red palm for two kinds of different generas that resistance is excellent, ornamental value is high are chosen respectively as male parent
And female parent；
2) artificial pollination：Through short-term 16 DEG C~18 DEG C stimulations or winter season, the pollen of male parent is gathered in florescence daystart, and
Artificial pollination is carried out to female parent, using the amber seed for being developed to maturation after pollination and not falling off as explant；
3) aseptic seeding culture：The outside rind of the amber seed is sloughed and cleaned up, is with concentration volume fraction first
Alcohol-pickled 30~40 seconds of 74%~77%, then sterilize 14 with the mercuric chloride solution that concentration mass fraction is 0.12%~0.18%
~16 minutes, then with rinsed with sterile water 5~7 times, finally blot the moisture of the amber seed outer surface with filter paper and connect
Kind, to be cultivated on 5.5~6.0 germination medium, is trained a height of 1~1.5 centimetre of plantlet to pH value；
The formula of germination medium is：1/2MS minimal mediums, activated carbon（AC）0.5~1.0g/L, 80~100mg/L of inositol,
1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.4mg/L, puridoxine hydrochloride (VB6) 0.4~1.0mg/L,
Nicotinic acid 0.3~0.6mg/L, 6- benzyl aminoadenine（6-BA）0.1~0.5mg/L, methyl α-naphthyl acetate（NAA）0.005~0.5 mg/L,
15~30g/L of sucrose, 4~5g/L of agar；
4) Multiplying culture：Step 3 will be passed through）In the plantlet that draws of culture to cut root and be transferred to pH values be 5.5~6.0
Cultivated in proliferated culture medium, be trained the seedling that height is 1.5~3.0 centimetres；
The formula of proliferated culture medium is：MS minimal mediums, 80~100mg/L of inositol, 1.0~2.0mg/L of glycine, hydrochloric acid
Thiamine (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4~1.0mg/L, 6- benzyl aminoadenine（6-BA）0.1
~1.0 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, 4~5g/L of agar；
5) strengthening seedling and rooting culture：Step 4 will be passed through）In the seedling that draws of culture be transferred to pH values and given birth to for 5.5~6.0 strong sprouts
Cultivated in root culture medium, be trained a height of 3.0~5.0cm test tube seedling；
The formula of strengthening seedling and rooting culture medium is：1/2MS minimal mediums, activated carbon（AC）0.5~3.0g/L, containing inositol 80~
100mg/L, 1.0~2.0mg/L of glycine, thiamine hydrochloride (VB1) 0.02~0.6mg/L, puridoxine hydrochloride (VB6) 0.4~
1.0mg/L, methyl α-naphthyl acetate（NAA）0.005~0.5 mg/L, 0.3~0.6mg/L of nicotinic acid, 20~30g/L of sucrose, 5~6g/ of agar
L；
6) test tube transplantation of seedlings：Step 5 will be passed through）In the test tube seedling that draws of culture be transferred to hardening in the greenhouse of shading more than 70%
5~10 days, then it is taken out from strengthening seedling and rooting culture medium, cleans the culture medium of root residual, with 70% thiophanate methyl or
Plant in seedling medium and cultivated into seedling after 800 times of liquid disinfectant processing of carbendazol wettable powder；
The formula of seedling medium is：It is 8 by volume by peat soil, perlite, river sand:1:1~9:0.5:0.5 is mixed to form
Mixed-matrix；
Step 2）In artificial pollination, when male parent is identical with the maternal florescence, maternal spadix gynoecium start secreting mucus 1~
In 2 days；Paternal pollen is obtained when daystart 4 points~6 directly to be invested in maternal column cap with writing brush；When male parent and maternal florescence not
During chance, take male parent material to be placed in phjytotron, wherein night sets 16 DEG C~18 DEG C of temperature, daytime 28 DEG C of growing temperatures~
32 DEG C, illuminance 2000lx, light irradiation time 12h/ d；Processing is more than two weeks, untill paternal pollen is collected, Zhi Houyong
Writing brush directly invests pollen in maternal column cap.
2. a kind of red palm crossbreeding and rapid propagation method as claimed in claim 1, it is characterised in that：It is characterized in that：Step
Rapid 3）~5）In cultivation temperature be 22~27 DEG C.
3. a kind of red palm crossbreeding and rapid propagation method as claimed in claim 1, it is characterised in that：Step 3）~5）In
Intensity of illumination is 1500~2000lx.
4. a kind of red palm crossbreeding and rapid propagation method as claimed in claim 1, it is characterised in that：Step 3）~5）Illumination
The h/d of Shi Changwei 10~14.
5. a kind of red palm crossbreeding and rapid propagation method as claimed in claim 1, it is characterised in that：Step 6）In illumination
Intensity is 1500~2500lx.
6. a kind of red palm crossbreeding and rapid propagation method as claimed in claim 1, it is characterised in that：The kind of the red palm
Can be Anthurium andraeanum（Great Ye fancy candles lit in the bridal chamber at wedding）、A.bakeri（Green spathe fancy candles lit in the bridal chamber at wedding）、A.crystallinum、
A.dussii (triangle leaf fancy candles lit in the bridal chamber at wedding), A.holtonianum（Anistree leaf fancy candles lit in the bridal chamber at wedding）、A.hookeri、A.kempteri、
A.longipetiolatum、A.macrolobum、A.magnificure、A.miquelianum、A.nymphillium、
A.ozfisianum、A.pedatoradiatum、A.putamayo、A.scanden、A.signature（Trilobated leaf fancy candles lit in the bridal chamber at wedding）、
A.subsignatum、A.undatum、A.variabile、A.veitchii、A.warocquenum。
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| CN110063258A (en) * | 2019-05-23 | 2019-07-30 | 三明市农业科学研究院 | A kind of red fruit beadplant quick breeding method for tissue culture |
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