CN105325297A - Seedling counting method for dendrobium officinale seeds - Google Patents
Seedling counting method for dendrobium officinale seeds Download PDFInfo
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- CN105325297A CN105325297A CN201510781009.5A CN201510781009A CN105325297A CN 105325297 A CN105325297 A CN 105325297A CN 201510781009 A CN201510781009 A CN 201510781009A CN 105325297 A CN105325297 A CN 105325297A
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- seeds
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- dendrobium candidum
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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
Abstract
The invention discloses a seedling counting method for dendrobium officinale seeds. The method comprises the steps that the dendrobium officinale seeds sprout to grow into seedlings, and the number of the seeds is obtained by calculating the number of the seedlings. According to the method, tiny dendrobium officinale capsule seeds which are difficult to directly distinguish and count with naked eyes by adopting a conventional method are sown on the surface of a culture medium, nutriment and water are provided for sprouting and growth of the dendrobium officinale seeds to ensure that the seeds preserved in the capsules can survive basically on the culture medium and sprout to grow into seedlings with a certain size capable of being clearly distinguished and counted with naked eyes, and counting can be visually, easily and conveniently achieved, so that the number of the seeds is obtained by counting the number of the seedlings. The method is easy and convenient to implement and high in practicability, the number of the seeds in the dendrobium officinale capsules can be accurately counted with out precision microscopic instruments so as to obtain the number of growth seedlings conveniently, and accurate guidance and reference are provided for large-scale seedling culture and production.
Description
Technical field
The invention belongs to plant trickle seed quick counter technical field, particularly relate to a kind of seedling counting method of Seeds of Dendrobium Candidum.
Background technology
Dendrobium candidum (DendrobiumofficinaleKimuraetMigo), the orchid family Dendrobium herbaceos perennial, 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 rate is not as good as 5% in its natural state for dendrobium candidum, and reproduction rate 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 seedling 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 problem: the seedling counting method of Seeds of Dendrobium Candidum, grows into seedling by the seed germination of dendrobium candidum, is obtained the quantity of seed by the quantity calculating seedling.
The seedling counting method of above-mentioned Seeds of Dendrobium Candidum, broadcasts the seed in dendrobium candidum capsule and makes its germination and growth become seedling in media surface.
Medium consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8.
The average plant height of seedling is more than or equal to 5.0mm.
The seedling counting method of above-mentioned Seeds of Dendrobium Candidum, comprises the following steps:
<1> medium preparing
Prepare medium 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 medium 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 seedling, average plant height reaches 5.0mm, takes out whole seedling 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 seedling counting method of Seeds of Dendrobium Candidum, the seed germination of dendrobium candidum is grown into seedling, is obtained the quantity of seed by the quantity calculating seedling.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 medium 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 medium (mature seed 99% is energy germination and growth on medium), and allow seed germination grow into naked eyes can know differentiate and counting a certain size seedling (average plant height reaches 5.0mm), make it directly perceived, easy counting, the quantity of seed then can be obtained by counting seedling 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
Medium consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8;
Prepare medium 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 medium 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, the first germination and growth of seed becomes protocorm, protocorm continues to cultivate and just grows up to the complete seedling of root, stem, leaf, until the seed culture of powdery after 100 days, the average plant height of seedling reaches 5.0mm (35.9 times for proembryo), and naked eyes can be easy to differentiate, and take out whole seedlings of each fruit, be one group of counting with 20 strains, be just easy to draw the grain weight in each capsule.
Result: the seed amount in 10 capsules is respectively 71500,43500,44500,26600,35196,53762,133100,95800,85300,75600.
Claims (5)
1. a seedling counting method for Seeds of Dendrobium Candidum, is characterized in that the seed germination of dendrobium candidum to grow into seedling, is obtained the quantity of seed by the quantity calculating seedling.
2. the seedling 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 seedling in media surface.
3. the seedling counting method of Seeds of Dendrobium Candidum according to claim 2, is characterized in that: described medium consists of MS+1.5mg/L6-BA+0.5mg/LNAA+200g/L potato leachate+20g/L sucrose+5g/L agar, pH5.8.
4. the seedling counting method of Seeds of Dendrobium Candidum according to claim 3, is characterized in that: the average plant height of described seedling is more than or equal to 5.0mm.
5. the seedling counting method of Seeds of Dendrobium Candidum according to claim 4, is characterized in that comprising the following steps:
<1> medium preparing
Prepare medium 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 medium 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 seedling, average plant height reaches 5.0mm, takes out whole seedling counting, obtains the quantity of seed in capsule.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510781009.5A CN105325297B (en) | 2015-11-13 | 2015-11-13 | The seedling counting method of Seeds of Dendrobium Candidum |
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| CN201510781009.5A CN105325297B (en) | 2015-11-13 | 2015-11-13 | The seedling counting method of Seeds of Dendrobium Candidum |
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| Publication Number | Publication Date |
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| CN105325297A true CN105325297A (en) | 2016-02-17 |
| CN105325297B CN105325297B (en) | 2018-07-17 |
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| CN201510781009.5A Expired - Fee Related CN105325297B (en) | 2015-11-13 | 2015-11-13 | The seedling counting method of Seeds of Dendrobium Candidum |
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Non-Patent Citations (3)
| 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》 * |
| 林江波等: "福建铁皮石斛人工繁育技术研究", 《福建农业学报》 * |
| 赵立子等: "柴胡种子质量分级标准", 《中国农学通报》 * |
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Granted publication date: 20180717 Termination date: 20191113 |