CN105469136A - Protocorm counting method of dendrobium candidum seeds - Google Patents
Protocorm counting method of dendrobium candidum seeds Download PDFInfo
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- CN105469136A CN105469136A CN201510781038.1A CN201510781038A CN105469136A CN 105469136 A CN105469136 A CN 105469136A CN 201510781038 A CN201510781038 A CN 201510781038A CN 105469136 A CN105469136 A CN 105469136A
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- protocorm
- seeds
- dendrobium candidum
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- counting
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- G—PHYSICS
- G06—COMPUTING; CALCULATING OR COUNTING
- G06M—COUNTING MECHANISMS; COUNTING OF OBJECTS NOT OTHERWISE PROVIDED FOR
- G06M11/00—Counting of objects distributed at random, e.g. on a surface
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
Abstract
The invention provides a protocorm counting method of dendrobium candidum seeds. Dendrobium candidum seeds are germinated into protocorms, and by counting the number of the protocorms, the number of the seeds is obtained. According to the method, the seeds, which are fine and difficult to count regularly and directly by eyes, in dendrobium candidum capsules are sown on the surface of a culture medium; abundant nutrition components of the culture medium are prepared, nutriments and water are provided for the germination and growth of the dendrobium candidum seeds, and the seeds retained by the capsules can basically survive on the culture medium; and the seeds are germinated into the certain-size protocorms which can be clearly distinguished and counted by eyes, and the protocorms can be visually and simply counted, and by counting the number of the protocorms, the number of the seeds is obtained. The method is convenient to use, is high in practicality and can accurately counting the number of seeds in the dendrobium candidum capsules without other relatively precise microscopic instruments, so that the number of production seedlings can be conveniently obtained, and accurate guidance and reference are provided for large-scale seeding culturing and production.
Description
Technical field
The invention belongs to plant trickle seed quick counter technical field, particularly relate to a kind of protocorm counting method of Seeds of Dendrobium Candidum.
Background technology
Dendrobium candidum (DendrobiumofficinaleKimuraetMigo), the orchid family Dendrobium per nnial herb, conventional Chinese medicine, by the Pharmacopoeia of the People's Republic of China is recorded.Be distributed in all provinces and regions on the south China Qinling Mountains, Huaihe River, grow on lofty mountains and steep hills overhanging cliff and rock crevice or remote mountains ancient tree.Dendrobium candidum have promote the production of body fluid, quench the thirst, effect of oedema of easing pain, eliminate, cure mainly the pyreticosis deficiency of Yin, order is dark, stomach is weak, the disease such as hoarseness, hoarseness tired to vocal cords, recovering its U.S. sound has special efficacy.Dendrobium candidum is endangered species, belongs to country-level national key protected plant, its fresh stem and frying dry after Tiepi Fengdou be all rare traditional Chinese medicine.Modern clinic application and pharmacological research confirm, dendrobium candidum has nourishing Yin and moistening lung, nourishing the stomach to improve the production of body fluid, brain nourishing and eyesight improving, the Proheparin heart, heat-clearing and fire-reducing, the consumptive disease of tonifying five zang organs, magical effect of prolonging life of making light of one's life by commiting suicide, and has definite curative effect and conditioning rehabilitative action to diseases such as chronic fatigue syndrome, tuberculosis, coronary heart disease, ephrosis, chronic liver disease, hypertension, high fat of blood, hyperglycaemia, diabetes, chronic gastropathy, dry syndrome, cancers; Strengthen body immunity, anti-oxidant, anti-ageing, to tobacco and wine excessively, overwork, night life is excessive, overstrain one's nerves with eye, hoarseness, mouth parched and tongue scorched, old gonosome low force decline, and the rehabilitations such as the deficiency of Yin have special efficacy.
Germination percentage is not as good as 5% in its natural state for dendrobium candidum, and breeding potential is very low, mainly with offshoot breeding, is difficult to meet the need of market.Present dendrobium candidum take mainly seed as explant, and the mode of carrying out tissue cultures production test-tube plantlet is bred.Seeds of Dendrobium Candidum is mainly present in capsule, capsule is about 1.8 ~ 2.2cm, diameter 0.5 ~ 0.8cm, interior containing several ten thousand to tens0000 seeds, tiny of dust-like, examine under a microscope kernel texture very simple, what only have a simple embryo and cell monolayer sticks up shape seed coat structure, without endosperm, and seed length × wide average out to 126 × 84 μm.Because the seed of dendrobium candidum is very trickle, be difficult to seed to separate, be difficult to grain weight contained by accurate counting capsule by conventional method, and seed not easily preserved in its natural state, be easy to inactivation.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of protocorm counting method of easy to use, practical Seeds of Dendrobium Candidum, with the seed amount in accurate counting dendrobium candidum capsule, instructs accurately and reference for growing dendrobium seedlings provides.
For solving the problems of the technologies described above, the present invention adopts following technical matters: the protocorm counting method of Seeds of Dendrobium Candidum, grows into protocorm by the seed germination of dendrobium candidum, is obtained the quantity of seed by the quantity calculating protocorm.
The protocorm counting method of above-mentioned Seeds of Dendrobium Candidum, broadcasts the seed in dendrobium candidum capsule and makes its germination and growth become protocorm in media surface.
Nutrient culture media consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8.
Protocorm diameter is more than or equal to 1mm.
The protocorm counting method of above-mentioned Seeds of Dendrobium Candidum, comprises the following steps:
<1> medium preparing
Prepare nutrient culture media by formula, and with the flat bottom glass bottle packing of diameter 6cm, high 10cm, every bottled 25ml, then puts 121 DEG C, 1.1kg/cm
2sterilized under pressure 20min, stand-by;
<2> Seed Treatment
Dendrobium candidum capsule uses liquid detergent aqueous solution soaking 5min, tap water 15-30min successively, then on superclean bench, puts into 1g/LHgCl
2sterilizing 15min in solution, after aseptic water washing 5 times, aseptic filter paper blots fruit surface moisture, stand-by;
<3> planting seed
By the seed access media surface in capsule;
<4> seed culture
The nutrient culture media connected is put in 25 DEG C of culturing room, supplementary illumination 10-12 hour/day, cultivate under intensity of illumination 1500lux condition;
<5> seed counting
Treat that seed grows up to protocorm, bulb diameter reaches 1mm, takes out whole protocorm counting, obtains the quantity of seed in capsule.
Seed for dendrobium candidum very trickle being difficult to is separated counting and the problem of not easily preserving, long period can be preserved in capsule according to seed, in production, Seed storage all adopts the mode that capsule is preserved, we have established a kind of protocorm counting method of Seeds of Dendrobium Candidum, the seed germination of dendrobium candidum is grown into protocorm, is obtained the quantity of seed by the quantity calculating protocorm.Because this genealogy of law is by trickle, be difficult to adopt conventional, seed in the dendrobium candidum capsule of direct discernable by eye counting is broadcast in media surface, utilize the nutrition that preparation nutrient culture media is abundant, sprout for Seeds of Dendrobium Candidum and grow and supply nutrients and moisture, guarantee that the seed that its capsule is preserved substantially can survive on nutrient culture media (mature seed 99% is energy germination and growth on nutrient culture media), and allow seed germination grow into naked eyes can know differentiate and counting a certain size protocorm (bulb diameter reaches 1mm), make it directly perceived, easy counting, the quantity of seed then can be obtained by counting protocorm quantity.The present invention is easy to use, practical, do not need by other more accurate microsurgical instrument equipment, just can seed amount in accurate counting dendrobium candidum capsule, thus conveniently drawing the quantity of producing seedling, this instructs and reference accurately for extensive nursery and production provide.
Embodiment
Embodiment
<1> medium preparing
Nutrient culture media consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8;
Prepare nutrient culture media by formula, and with the flat bottom glass bottle packing of diameter 6cm, high 10cm, every bottled 25ml, then puts 121 DEG C, 1.1kg/cm
2sterilized under pressure 20min, stand-by;
<2> Seed Treatment
Late September is to late November, gathering, healthy full, free from insect pests, pericarp slightly turn yellow, the ripe dendrobium candidum capsule that do not ftracture uses liquid detergent aqueous solution soaking 5min, tap water 15-30min successively, remove unnecessary carpopodium, base of fruit etc., then on superclean bench, put into 1g/LHgCl
2sterilizing 15min in solution, after aseptic water washing 5 times, aseptic filter paper blots fruit surface moisture, stand-by;
<3> planting seed
At superclean bench, in end-grain cutting one osculum of capsule, shake capsule lightly, the seed in capsule is accessed media surface thinly, equably; Sow 10 capsules altogether, each fruit one numbering, according to fruit size, every carpostrote tens bottles to more than hundred bottles is not etc.;
<4> seed culture
The nutrient culture media connected is put in 25 DEG C of culturing room, supplementary illumination 10-12 hour/day, cultivate under intensity of illumination 1500lux condition;
<5> seed counting
Carrying out in dendrobium officinale test-tube plantlet incubation, seed germination grows into the protocorm that color and luster is dark green, sparkling and crystal-clear, have phyllopodium, until the seed culture of powdery after 60 days, the length and width of protocorm reach 1592 μm × 939 μm (1.6mm × 0.9mm, length and width increase 11.9 times and 10.7 times respectively than proembryo), naked eyes can be easy to differentiate, and take out whole protocorms of each fruit, be one group of counting with 20, be just easy to draw the grain weight in each capsule.
Result: the seed amount in 10 capsules is respectively 71500,42500,45600,27600,35196,54762,123100,96850,75680,43560.
Further, the complete seedling of root, stem, leaf is just grown up to because protocorm continues to cultivate, just cannot continue to sprout if protocorm diameter does not reach 1mm, simultaneously in conjunction with the growthform of protocorm, just accurately can judge that it is with or without the ability developing into seedling further, thus infer the developmental potency of seed in a capsule.Therefore, application the present invention both can know and calculated seed amount in capsule, can accurately judge again seed survival in capsule quantity (protocorm diameter be more than or equal to 1mm just for viable seeds quantity, be less than 1mm for dead seed amount), thus provide Data support more accurately for production and selling.
Claims (5)
1. a protocorm counting method for Seeds of Dendrobium Candidum, is characterized in that the seed germination of dendrobium candidum to grow into protocorm, is obtained the quantity of seed by the quantity calculating protocorm.
2. the protocorm counting method of Seeds of Dendrobium Candidum according to claim 1, is characterized in that the seed in dendrobium candidum capsule to broadcast to make its germination and growth become protocorm in media surface.
3. the protocorm counting method of Seeds of Dendrobium Candidum according to claim 2, is characterized in that: described nutrient culture media consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8.
4. the protocorm counting method of Seeds of Dendrobium Candidum according to claim 3, is characterized in that: described protocorm mean diameter is more than or equal to 1mm.
5. the protocorm counting method of Seeds of Dendrobium Candidum according to claim 4, is characterized in that comprising the following steps:
<1> medium preparing
Prepare nutrient culture media by formula, and with the flat bottom glass bottle packing of diameter 6cm, high 10cm, every bottled 25ml, then puts 121 DEG C, 1.1kg/ ㎝
2sterilized under pressure 20min, stand-by;
<2> Seed Treatment
Dendrobium candidum capsule uses liquid detergent aqueous solution soaking 5min, tap water 15-30min successively, then on superclean bench, puts into 1g/LHgCl
2sterilizing 15min in solution, after aseptic water washing 5 times, aseptic filter paper blots fruit surface moisture, stand-by;
<3> planting seed
By the seed access media surface in capsule;
<4> seed culture
The nutrient culture media connected is put in 25 DEG C of culturing room, supplementary illumination 10-12 hour/day, cultivate under intensity of illumination 1500lux condition;
<5> seed counting
Treat that seed grows up to protocorm, average bulb diameter reaches 1mm, takes out whole protocorm counting, obtains the quantity of seed in capsule.
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| CN201510781038.1A CN105469136B (en) | 2015-11-13 | 2015-11-13 | The protocorm counting method of Seeds of Dendrobium Candidum |
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907323A (en) * | 2012-11-01 | 2013-02-06 | 建始县颐泰生物科技有限公司 | Method for aseptically producing miniature seed stems of common bletilla pseudobulb seeds |
| CN103392593A (en) * | 2013-06-20 | 2013-11-20 | 武汉久源生物医药科技有限公司 | Method for rapid tissue culture propagation of Dendrobium huoshanense seedlings |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102907323A (en) * | 2012-11-01 | 2013-02-06 | 建始县颐泰生物科技有限公司 | Method for aseptically producing miniature seed stems of common bletilla pseudobulb seeds |
| CN103392593A (en) * | 2013-06-20 | 2013-11-20 | 武汉久源生物医药科技有限公司 | Method for rapid tissue culture propagation of Dendrobium huoshanense seedlings |
Non-Patent Citations (1)
| Title |
|---|
| 付传明等: "《铁皮石斛无菌播种产业化繁育技术研究》", 《广西植物》 * |
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