CN102144536A - Method for crossbreeding and aseptic sowing of renanthera coccinea - Google Patents
Method for crossbreeding and aseptic sowing of renanthera coccinea Download PDFInfo
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Abstract
The invention discloses a method for crossbreeding and aseptic sowing of renanthera coccinea, comprising the following steps of: carrying out hybridization by selecting orchid with obvious characters and high visual value as a hybrid female parent and renanthera coccinea as a male parent; carrying out aseptic sowing on a seed germination medium on the fruit obtained by successful pollination when being basically ripened but not splitted; after germinating, transferring the seeds to a healthy seedling medium; and growing the formed seedling in a test tube out of a bottle after being hardened under the illumination of natural light for 10 to 20 days, wherein the transplanting survival rate of seedling formation can reach 90% or more. The method has the characteristic of high hybrid success rate, and is particularly useful during taking orchid with obvious characters and high visual value as a hybrid male parent which is different from a renanthera coccinea parent in flowering phase. The unique medium and cultivation method are employed by the invention so that the hybrid seeds of renanthera coccinea have high germination, fast seedling formation and good seedling quality. The method has the advantages of strong operation, effective breeding, valuable application and the like.
Description
Technical field:
The present invention relates to the cross breeding method of plant, be specifically related to a kind of crossbreeding and aseptic seeding method of Herba Renantherae coccineae.
Background technology:
Flame Cymbidium (Reanathera) the plant whole world has 21 kinds approximately, for growing nonparasitically upon another plant or semiepiphyte, be distributed in Southeast Asia to the Himalaya area, comprise the South China, China originates in 3 kinds, and wherein Herba Renantherae coccineae (Ren.Coccinea) originates in Guangdong, area, Hainan, the tall and big growth of plant is healthy and vigorous, and Yunnan Herba Renantherae coccineae (Ren.imschootiana) and Chinese Herba Renantherae coccineae (Ren.sinica) originate in Yunnan.Herba Renantherae coccineae growth is healthy and vigorous, and pattern is gorgeous, spends number numerous, far away see as flame as gorgeous, have that very high horticultural applications is worth and breeding value, be top grade flower all the fashion on the International Flower market, have a large amount of fans in the world.And, be adapted at South China's cultivation because its growing environment is the torrid zone, subtropics, and be hopeful future to set up its breeding and production base in the South China, form the industry of similar Moth orchid, dendrobium.Herba Renantherae coccineae has become one of plant species in imminent danger in the world, and all wild kinds all are put into " CITS " (CITES) in the appendix and the trade that is under an embargo.
The Herba Renantherae coccineae crossbreed is more more graceful than the colored type flower of initial species appearance, growth potential and strong stress resistance, more gorgeous more colorful than initial species on pattern, also can conclude the business in the international market, now become the main product of world's Herba Renantherae coccineae industry, but the crossbreeding work of China's Herba Renantherae coccineae is also at the early-stage, and the Herba Renantherae coccineae industry has vast market prospect.
Though Herba Renantherae coccineae has very high cross compatibility, but with other orchid hybridization the time, there is the flowering asynchronism problem, can't carry out some distant hybridization owing to flowering asynchronism simultaneously, the pollen for low temperature storage of the other plant of having reported is in addition gone mouldy problem often and is caused pollen to preserve failure.The Herba Renantherae coccineae hybrid seed is owing to its embryo ateliosis, and germination rate is extremely low under the nature, needs aseptic seeding just can carry out seedling production.
Summary of the invention:
The purpose of this invention is to provide a kind of crossbreeding of Herba Renantherae coccineae method and germination rate height, become that seedling is fast, the method for the aseptic seeding of seedling quality better.
The present invention selects the high orchid of the obvious characteristic of tool, ornamental value maternal as hybridization, with Herba Renantherae coccineae as male parent, hybridize, when the fruit that successfully obtains of pollinating does not ftracture being mature on the whole, carry out aseptic seeding on the seed germination medium, transfer on the strong seedling culture base behind the seed germination again and cultivate, the test-tube plantlet of formation is through the bottle outlet cultivation after 10～20 days of natural lighting lower refining seedling, the transplanting survival rate of Cheng Miao can reach more than 90%, thereby has realized purpose of the present invention.
The crossbreeding of Herba Renantherae coccineae of the present invention and the method for aseptic seeding is characterized in that, may further comprise the steps:
(1): artificial pollination:, as male parent, gather the pollen of male parent and invest on the maternal column cap, and carry out bagging and handle as female parent with orchid, prevent the interference of foreign pollen with Herba Renantherae coccineae;
(2): aseptic seeding: the hybridization capsule of bearing when female parent is grown and is mature on the whole and fruit when not ftractureing, this fruit put on the skin wipe clean, be after 70～75% alcoholic solutions soak 2～3 minutes with volume fraction successively, placing mass fraction is that 0.1%～0.2% mercuric chloride solution was sterilized 20～30 minutes, again with sterile water towards Xian 4～5 times, blot the moisture of fruit surface, cut fruit, Powdered seed is inoculated on the seed germination medium cultivates, seed germination forms protocorm earlier, grow up to seedling subsequently, described seed germination medium is every liter and contains and spend precious No. 1 1～3.0g, peptone 0.5～2.0g, coconut milk 100～150mL, sucrose 20～30g, agar 6～7g, active carbon 0.5～1.0g, inositol 80～120mg, glycine 1.5～2.5mg, thiamine hydrochloride 0.05～0.2mg, puridoxine hydrochloride 0.4～0.8mg, nicotinic acid 0.4～0.8mg, surplus is a water, and pH 5.4～5.8; Be inoculated on the seed germination medium from seed and cultivate, seed germination formed protocorm in general 30～90 days, further formed seedling, seed germination rate 60～90%, planting percent 55～90% in 90～180 days;
(3): strong seedling culture: the seedling that aseptic seeding is obtained is inoculated on the strong seedling culture base to be cultivated, can form healthy and strong plant in the time of 120～180 days, described strong seedling culture base is every liter and contains and spend precious No. 1 1～3.0g, peptone 0.5～2.0g, methyl 0.5～1.0mg, banana homogenate 50～100g, coconut milk 50～100mL, sucrose 15～20g, agar 6～7g, active carbon 0.5～2.0g, inositol 80～120mg, glycine 1.5～2.5mg, thiamine hydrochloride 0.05～0.2mg, puridoxine hydrochloride 0.4～0.8mg, nicotinic acid 0.4～0.8mg, surplus is a water, pH 5.4～5.8;
(4): test-tube seedling transplanting: when the plant 5～8cm that grows up to when seedling is high, blake bottle was transferred to the natural daylight lower refining seedling 10～20 days, then it is taken out, clean the medium of root, move into blue stone: wood chip: bark is 2～3: 1～2 by volume: cultivate in 1～2 the mixed-matrix; Routine Management keeps suitable ventilation and enough humidity, and the survival rate of transplanting can reach more than 90%;
Condition of culture in above-mentioned steps (2) and (3) is: 24～28 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12～16 hours/day.
In described artificial pollination step, when female parent is identical with the male parent florescence, launched the back 2～4 days at maternal flower, get the pollen of male parent and directly invest on the maternal column cap.
In described artificial pollination step, when the flowering asynchronism of maternal and male parent, full-bloom stage at the male parent Herba Renantherae coccineae is gathered pollinium, gather choose opportunities when Herba Renantherae coccineae flower petal has just launched, acquisition time is chosen in the morning 8～9 o'clock, gather pollen and place aseptic plastic tube, sealing places 4 ℃ to keep in Dark Place, when female parent is bloomed, the paternal pollen that low temperature the is preserved 20～30min that under 15～35 ℃ condition, thaws, add sterile water in the plastic tube of placing pollen again and soak back wet pollen 20～30min, then viable pollen is invested on the maternal column cap.Carry out the preservation of the pollen of Herba Renantherae coccineae according to the method described above, the holding time of Herba Renantherae coccineae pollen can reach 3～10 months.The test of pollen viability can adopt chlorinated triphenyl tetrazole decoration method to measure pollen viability, confirms after pollen has vigor it to be invested on the maternal column cap.The above-mentioned clean low temperature that adopts is preserved has before blodynamic Herba Renantherae coccineae pollen, the pollination pollen thaw the problem of going mouldy when technology such as returning wet process has solved that pollen is preserved and the hybridization pollination obstruction of the kind at different florescences and plant, thereby lays the first stone for the Herba Renantherae coccineae breeding of new variety.
Described collection pollen places aseptic plastic tube, sealing places 4 ℃ to keep in Dark Place, preferably, step is: use aseptic toothpick and choose colored anther cap gently, with aseptic toothpick pollinium is sticked on the toothpick point and also rapidly pollinium is put to the 1ml centrifuge tube of cleaning sterile, every pipe is put into the pollinium of 1 flower, will place the centrifuge tube of pollinium and concentrate dress with an aseptic vial again, and good seal, place 4 ℃ to keep in Dark Place.
The maturing stage difference of different Herba Renantherae coccineae crossbreed fruits after pollination, but most of Herba Renantherae coccineae hybrid seed just can be sprouted in the time of back 4 months in pollination, reach during by 5～6 months and sprout the peak, descend then, but need 10～12 months with the fruit development of the Herba Renantherae coccineae hybrid of Cymbidium hybridization is ripe.The germination period of general Herba Renantherae coccineae hybrid seed the best is to be mature on the whole and fruit when not ftractureing at fruit development.
Aseptic seeding step in step (2), preferably the protocorm that seed germination is formed earlier is inoculated on the differential medium, make protocorm on differential medium, form seedling, described differential medium is every liter of interpolation 6-Bian Ji adenine 5.0mg on the basis of seed germination medium, condition of culture is: 24～28 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12～16 hours/day.
Described maternal orchid is preferably: Doritis pulcherrima (Doritis pulcherrima), Xiao Hua all ages blue (Vandacoerulescens) or color heart sword-leaved cymbidium (Cymbidium ensifolium).
The treasured of spending that uses in step (2) and (3) medium is commercially available for No. 1.
The present invention has hybridization success rate high characteristics, and is very effective when particularly the high orchid of the obvious characteristic of tool, the ornamental value of some and the flowering asynchronism of Herba Renantherae coccineae hybrid strain being hybrid strain.The present invention adopts unique medium and cultural method, make the Herba Renantherae coccineae hybrid seed the germination rate height, become that seedling is fast, the seedling quality better.The present invention has strong operability, breeding efficiency and using value advantages of higher.
Embodiment:
Below be to further specify to of the present invention, rather than limitation of the present invention.
That uses among the embodiment spends precious No. 1 (HYPONeX 1) for produced in USA, Taiwan platform and gardening enterprise stock action Co., Ltd packing product.
Embodiment 1:
The crossbreeding and the sapling multiplication of Doritis pulcherrima (Doritis pulcherrima) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana)
(1) artificial pollination: the Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, when just having launched, gathers on its single flower petal pollen, o'clock use sterilized toothpick in the morning 8～9 and choose colored anther cap gently, with toothpick pollinium is sticked on the toothpick point again and also rapidly pollinium is put to the 1ml EP pipe (centrifuge tube) of cleaning sterile, every pipe is put into the pollinium of 1 flower, concentrate dress to get up and good seal with an aseptic vial again with having placed pollinium EP pipe, the vial that will fill pollen at last is positioned under 4 ℃ of conditions and keeps in Dark Place.After the time that Herba Renantherae coccineae pollen is preserved reaches 3 months, when maternal Doritis pulcherrima when blooming at the beginning of 6 months then, the pollen of the Herba Renantherae coccineae that low temperature is preserved thaws and returns wet process, to recover and the vitality of raising pollen.The first Herba Renantherae coccineae pollen that low temperature is preserved after thawing 20 minutes under room temperature (35 ℃) condition, add several sterile waters in the EP pipe of placing pollinium again and returned wet process 20 minutes, adopt chlorinated triphenyl tetrazole decoration method to measure pollen viability at last, determine that pollen has vigor, it is invested on the maternal column cap.Bagging behind the lip of pollination back excision female parent removes bag after about 1 week, hang up label, has information such as father, maternal title, hybridization date and pollination people on the label.Note to observe its developmental state, the success rate of statistics pollination in the time of 3 months, ripening rate all reaches more than 90%, and it is orange-yellow to wait fruit to become, and reaches to be mature on the whole and can to gather when not ftractureing and carry out aseptic seeding.
(2) aseptic seeding: the fruit that will pollinate back 5 months is put on the skin with the alcohol cotton earlier and is wiped clean the fruit surface dirt, be after 70% alcoholic solution soaks 2 minutes then with volume fraction, placing mass fraction is 0.1% mercuric chloride solution sterilization 30 minutes, after the rinsed with sterile water 5 times, blot the moisture of fruit surface with aseptic filter paper, cut fruit, Powdered embryo is inoculated on the seed germination medium, the sterilization success rate is more than 95%, visible seed expands in the time of 20 days changes green, seed germination forms protocorm in the time of 30 days, 90 days further seedlings that form.Seed germination rate 90%, planting percent 90%.The seed germination medium is every liter and contains and spend precious No. 1 2.0g, peptone 2.0g, coconut milk 100mL, sucrose 20g, agar 6g, active carbon 1.0g, inositol 80mg, glycine 2.5mg, thiamine hydrochloride 0.05mg, puridoxine hydrochloride 0.8mg, nicotinic acid 0.4mg, surplus is a water, pH 5.8.
(3) strong seedling culture: the seedling that aseptic seeding is obtained is inoculated on the strong seedling culture base to be cultivated, and can form the plant of stalwartness in the time of 120 days.Described strong seedling culture base is every liter and contains and spend precious No. 1 3.0g, peptone 2.0g, methyl 1mg, banana homogenate 80g, coconut milk 50mL, sucrose 15g, agar 6g, active carbon 2.0g, inositol 80mg, glycine 2.5mg, thiamine hydrochloride 0.05mg, puridoxine hydrochloride 0.8mg, nicotinic acid 0.4mg, surplus is a water, pH 5.8.
(4) test-tube seedling transplanting: in the time of the about 5-8 of plant centimetre high, blake bottle was transferred to the natural daylight lower refining seedling 15 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark is in 2: 1: 1 the mixed-matrix by volume, Routine Management keeps suitably ventilating and enough humidity, and the survival rate of transplanting all can reach more than 95%.
Condition of culture in step (2) and step (3) is 24 ℃ of cultivation temperature, illuminance 2000lx, illumination 12 hours/day.
Embodiment 2:
The crossbreeding and the sapling multiplication of Xiao Hua all ages blue (Vanda coerulescens) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana)
(1) artificial pollination: the Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, when just having launched, gathers on its single flower petal pollen, o'clock use sterilized toothpick in the morning 8～9 and choose colored anther cap gently, with aseptic toothpick pollinium is sticked on the toothpick point again and also rapidly pollinium is put to the 1mlEP pipe of cleaning sterile, every pipe is put into the pollinium of 1 flower, concentrate dress to get up and good seal with an aseptic vial again with having placed pollinium EP pipe, the vial that will fill pollen at last is positioned under 4 ℃ of conditions and keeps in Dark Place.After the time that Herba Renantherae coccineae pollen is preserved reached 6 months, all ages blue when blooming at the beginning of 9 months then as maternal Xiao Hua, the pollen of the Herba Renantherae coccineae that low temperature is preserved thawed and returns wet process, to recover and the vitality of raising pollen.Earlier the Herba Renantherae coccineae pollen that low temperature is preserved adds several sterile waters in the EP pipe of placement pollinium again after (30 ℃) thawed 25 minutes at ambient temperature and returned wet process 25 minutes, adopt chlorinated triphenyl tetrazole decoration method to measure pollen viability at last, determine that pollen has vigor, it is invested on the maternal column cap.Bagging behind the lip of pollination back excision female parent removes bag after about 1 week, hang up label, has information such as father, maternal title, hybridization date and pollination people on the label.Note observing its developmental state, the success rate of statistics pollination in the time of 3 months, ripening rate all reaches more than 80%, and it is orange-yellow to wait fruit to become, and is mature on the whole can gather when not ftractureing to carry out aseptic seeding.
(2) aseptic seeding: the fruit that will pollinate back 5 months is put on the skin with the alcohol cotton earlier and is wiped clean the fruit surface dirt, be 75% after alcohol-pickled 3 minutes then with volume fraction, placing mass fraction is 0.2% mercuric chloride solution sterilization 20 minutes, after the rinsed with sterile water 4 times, blot the moisture that fruit shows with aseptic filter paper, cut fruit, Powdered embryo is inoculated on the seed germination medium, the sterilization success rate reaches 100%, visible seed germination in the time of 30 days, seed germination forms protocorm in the time of 60 days, 120 days further seedlings that form.Seed germination rate 80%, planting percent 60%.The seed germination medium is every liter and contains and spend precious No. 1 3.0g, peptone 1.5g, coconut milk 150mL, sucrose 30g, agar 7g, active carbon 0.5g, inositol 120mg, glycine 1.5mg, thiamine hydrochloride 0.2mg, puridoxine hydrochloride 0.4mg, nicotinic acid 0.8mg, surplus is a water, pH 5.4.
(3) strong seedling culture: the seedling that aseptic seeding is obtained is inoculated on the strong seedling culture base to be cultivated, and can form the plant of stalwartness in the time of 150 days.Described strong seedling culture base is every liter and contains and spend precious No. 1 2.0g, peptone 1.5g, methyl 0.5mg, banana homogenate 100g, coconut milk 100mL, sucrose 20g, agar 7g, active carbon 1.0g, inositol 120mg, glycine 1.5mg, thiamine hydrochloride 0.2mg, puridoxine hydrochloride 0.4mg, nicotinic acid 0.8mg, surplus is a water, pH 5.4.
(4) test-tube seedling transplanting: when about 5～8 centimetres high of plant, blake bottle was transferred to the natural daylight lower refining seedling 20 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark is in 1: 1: 1 the mixed-matrix by volume, Routine Management keeps suitably ventilating and enough humidity, and the survival rate of transplanting all can reach more than 90%.
The condition of culture of step (2) and (3) is 28 ℃ of cultivation temperature, illuminance 1500lx, illumination 14 hours/day.
Embodiment 3:
The crossbreeding and the sapling multiplication of color heart sword-leaved cymbidium (Cymbidium ensifolium) and Yunnan Herba Renantherae coccineae (Renanthera imschootiana)
(1) artificial pollination: the Yunnan Herba Renantherae coccineae was bloomed at the beginning of 3 months, when just having launched, gathers on its single flower petal pollen, o'clock use sterilized toothpick in the morning 8～9 and choose colored anther cap gently, with aseptic toothpick pollinium is sticked on the toothpick point again and also rapidly pollinium is put to the 1mlEP pipe of cleaning sterile, every pipe is put into the pollinium of 1 flower, concentrate dress to get up and good seal with an aseptic vial again with having placed pollinium EP pipe, the vial that will fill pollen at last is positioned under 4 ℃ of conditions and keeps in Dark Place.After the time that Herba Renantherae coccineae pollen is preserved reaches 9 months, when the color heart sword-leaved cymbidium of female parent when blooming at the beginning of 12 months then, the pollen of the Herba Renantherae coccineae that low temperature is preserved thaws and returns wet process, to recover and the vitality of raising pollen.The Herba Renantherae coccineae pollen that elder generation preserves low temperature, after thawing 30 minutes under room temperature (15 ℃) condition, add several sterile waters in the EP pipe of placing pollinium again and returned wet process 30 minutes, adopt chlorinated triphenyl tetrazole decoration method to measure pollen viability at last, determine that pollen has vigor, it is invested on the maternal column cap.Bagging behind the lip of pollination back excision female parent removes bag after about 1 week, hang up label, has information such as father, maternal title, hybridization date and pollination people on the label.Note observing its developmental state, the success rate of statistics pollination in the time of 3 months, ripening rate reaches 30%, and it is orange-yellow to wait fruit to become, and is mature on the whole can gather when not ftractureing to carry out aseptic seeding.
(2) aseptic seeding: the fruit that will pollinate back 12 months is put on the skin with the alcohol cotton earlier and is wiped clean the fruit surface dirt, be 70% after alcohol-pickled 2.5 minutes then with volume fraction, placing mass fraction is 0.2% mercuric chloride solution sterilization 25 minutes, after the rinsed with sterile water 5 times, blot the water of fruit surface with aseptic filter paper, cut fruit, Powdered embryo is inoculated on the seed germination medium, the sterilization success rate reaches 100%, visible seed germination in the time of 90 days, seed germination forms root-like stock in the time of 120 days, and the root-like stock inoculated and cultured in differential medium, can be differentiated to form seedling in 60 days.Seed germination rate 60%, planting percent 55%.The seed germination medium contains for every liter spends precious No. 1 1g, peptone 0.5g, coconut milk 120mL, sucrose 25g, agar 6.5g, active carbon 0.7g, inositol 100mg, glycine 2mg, thiamine hydrochloride 0.1mg, puridoxine hydrochloride 0.5mg, nicotinic acid 0.5mg, surplus is a water, pH 5.6.Differential medium is every liter of interpolation 6-Bian Ji adenine 5.0mg on the basis of seed germination medium.
(3) strong seedling culture: the seedling that root-like stock differentiation is obtained is inoculated on the strong seedling culture base to be cultivated, and can form the plant of stalwartness in the time of 150 days.Described strong seedling culture base is every liter and contains and spend precious No. 1 1.0g, peptone 0.5g, methyl 0.7mg, banana homogenate 50g, coconut milk 75mL, sucrose 18g, agar 6.5g, active carbon 0.5g, inositol 100mg, glycine 2mg, thiamine hydrochloride 0.1mg, puridoxine hydrochloride 0.5mg, nicotinic acid 0.5mg, surplus is a water, pH 5.6.
(4) test-tube seedling transplanting: when about 5～8 centimetres high of plant, blake bottle was transferred to the natural daylight lower refining seedling 10 days, then it is taken out from vial, clean the medium of root, move into blue stone: wood chip: bark is in 3: 1: 2 the mixed-matrix by volume, Routine Management keeps suitably ventilating and enough humidity, and the survival rate of transplanting all can reach more than 95%.
Step (2) is cultivated in seed germination medium and differential medium and the condition of culture of step (3) is 26 ℃ of cultivation temperature, illuminance 1800lx, illumination 16 hours/day.
Claims (6)
1. the method for the crossbreeding of a Herba Renantherae coccineae and aseptic seeding is characterized in that, may further comprise the steps:
(1): artificial pollination: as female parent, as male parent, gather the pollen of male parent and invest on the maternal column cap, and carry out bagging and handle with Herba Renantherae coccineae with orchid;
(2): aseptic seeding: the hybridization capsule of bearing when female parent is grown and is mature on the whole and fruit when not ftractureing, this fruit put on the skin wipe clean, be after 70～75% alcoholic solutions soak 2～3 minutes with volume fraction successively, placing mass fraction is that 0.1%～0.2% mercuric chloride solution was sterilized 20～30 minutes, again with sterile water towards Xian 4～5 times, blot the moisture of fruit surface, cut fruit, Powdered seed is inoculated on the seed germination medium cultivates, seed germination forms protocorm earlier, grow up to seedling subsequently, described seed germination medium is every liter and contains and spend precious No. 1 1～3.0g, peptone 0.5～2.0g, coconut milk 100～150mL, sucrose 20～30g, agar 6～7g, active carbon 0.5～1.0g, inositol 80～120mg, glycine 1.5～2.5mg, thiamine hydrochloride 0.05～0.2mg, puridoxine hydrochloride 0.4～0.8mg, nicotinic acid 0.4～0.8mg, surplus is a water, and pH 5.4～5.8;
(3): strong seedling culture: the seedling that aseptic seeding is obtained is inoculated on the strong seedling culture base to be cultivated, described strong seedling culture base is every liter and contains and spend precious No. 1 1～3.0g, peptone 0.5～2.0g, methyl 0.5～1.0mg, banana homogenate 50～100g, coconut milk 50～100mL, sucrose 15～20g, agar 6～7g, active carbon 0.5～2.0g, inositol 80～120mg, glycine 1.5～2.5mg, thiamine hydrochloride 0.05～0.2mg, puridoxine hydrochloride 0.4～0.8mg, nicotinic acid 0.4～0.8mg, surplus is a water, and pH 5.4～5.8;
(4): test-tube seedling transplanting: when the plant 5～8cm that grows up to when seedling is high, blake bottle was transferred to the natural daylight lower refining seedling 10～20 days, then it is taken out, clean the medium of root, move into blue stone: wood chip: bark is 2～3: 1～2 by volume: cultivate in 1～2 the mixed-matrix;
Condition of culture in above-mentioned steps (2) and (3) is: 24～28 ℃ of cultivation temperature, illuminance 1500～2000lx, illumination 12～16 hours/day.
2. the method for the crossbreeding of Herba Renantherae coccineae according to claim 1 and aseptic seeding, it is characterized in that, in described artificial pollination step, when female parent is identical with the male parent florescence, launched the back 2～4 days at maternal flower, get the pollen of male parent and directly invest on the maternal column cap.
3. the method for the crossbreeding of Herba Renantherae coccineae according to claim 1 and aseptic seeding, it is characterized in that, in described artificial pollination step, when the flowering asynchronism of maternal and male parent, full-bloom stage at the male parent Herba Renantherae coccineae is gathered pollinium, gather choose opportunities when Herba Renantherae coccineae flower petal has just launched, acquisition time is chosen in the morning 8～9 o'clock, gather pollen and place aseptic plastic tube, sealing places 4 ℃ to keep in Dark Place, when female parent is bloomed, the paternal pollen that low temperature the is preserved 20～30min that under 15～35 ℃ condition, thaws, add sterile water in the plastic tube of placing pollen again and soak back wet pollen 20～30min, then viable pollen is invested on the maternal column cap.
4. the method for the crossbreeding of Herba Renantherae coccineae according to claim 3 and aseptic seeding, it is characterized in that, described collection pollen places aseptic plastic tube, sealing places 4 ℃ to keep in Dark Place, step is: use aseptic toothpick and choose colored anther cap gently, with aseptic toothpick pollinium is sticked on the toothpick point again and also rapidly pollinium is put to the 1ml centrifuge tube of cleaning sterile, every pipe is put into the pollinium of 1 flower, the centrifuge tube of having placed pollinium is concentrated dress with an aseptic vial again, and good seal, place 4 ℃ to keep in Dark Place.
5. the method for the crossbreeding of Herba Renantherae coccineae according to claim 1 and aseptic seeding, it is characterized in that, aseptic seeding step in step (2), the protocorm that seed germination is formed earlier is inoculated on the differential medium, make protocorm form seedling on differential medium, described differential medium is every liter of interpolation 6-Bian Ji adenine 5.0mg on the basis of seed germination medium, and condition of culture is: 24～28 ℃ of temperature, illuminance 1500～2000lx, illumination 12～16 hours/day.
6. the method for the crossbreeding of Herba Renantherae coccineae according to claim 1 and aseptic seeding, it is characterized in that described maternal orchid is: Doritis pulcherrima (Doritis pulcherrima), Xiao Hua all ages blue (Vanda coerulescens) or color heart sword-leaved cymbidium (Cymbidium ensifolium).
Priority Applications (1)
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106172005A (en) * | 2016-08-03 | 2016-12-07 | 济南麒麟花卉有限公司 | A kind of subculture medium and preparation method thereof |
| CN106489730A (en) * | 2016-09-30 | 2017-03-15 | 中国科学院华南植物园 | A kind of Herba Renantherae coccineae once-seedling forming quick propagating method and culture medium |
| CN109122302A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
| CN109122301A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of Doritis pulcherrima and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
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| CN1586167A (en) * | 2004-07-30 | 2005-03-02 | 中国科学院华南植物园 | Quick breeding technolgy for Renanthera imschootiana Rolfe |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106172005A (en) * | 2016-08-03 | 2016-12-07 | 济南麒麟花卉有限公司 | A kind of subculture medium and preparation method thereof |
| CN106489730A (en) * | 2016-09-30 | 2017-03-15 | 中国科学院华南植物园 | A kind of Herba Renantherae coccineae once-seedling forming quick propagating method and culture medium |
| CN106489730B (en) * | 2016-09-30 | 2018-09-28 | 中国科学院华南植物园 | A kind of Herba Renantherae coccineae once-seedling forming quick propagating method and culture medium |
| CN109122302A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of sword-leaved cymbidium and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
| CN109122301A (en) * | 2018-10-19 | 2019-01-04 | 中国科学院华南植物园 | A kind of method of Doritis pulcherrima and Herba Renantherae coccineae generic cross cultivation orchid new varieties |
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