CN103348919B - Dendrobe breeding method - Google Patents
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- CN103348919B CN103348919B CN201310314821.8A CN201310314821A CN103348919B CN 103348919 B CN103348919 B CN 103348919B CN 201310314821 A CN201310314821 A CN 201310314821A CN 103348919 B CN103348919 B CN 103348919B
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Abstract
The invention provides a dendrobe breeding method, and relates to a plant variety breeding method. The dendrobe breeding method comprises the following steps: collecting Dendrobium officinale seeds, selfing parent plants gained through dendrobe seed breeding to gain capsule, cultivating the capsule to gain germchit, screening better germchit, and hardening-seedling and transplanting after asexual reproduction of the germchit. Through the hybridization to the Dendrobium officinale parent plants, the method of combining a sexual reproduction manner and an asexual reproduction manner is adopted to breed a dendrobe clone variety; the dendrobe clone variety is suitable for being planted in a plurality of places in Yunnan such as Wenshan, Banna, Puer, Baoshan, Dehong and the like; an important value is brought to improve the mess dendrobe planting varieties in Yunnan province, enrich dendrobe seed resource and push seed selection adapting to local condition by peasant households.
Description
Technical field
The present invention relates to a kind of plant variety breeding method, particularly relate to a kind of breeding method of the stem of noble dendrobium.
Background technology
Dendrobium candidum
dendrobium officinalekimura et Migo belongs to the orchid family perennial herbaceous plant that grows nonparasitically upon another plant, and in motherland's traditional medicine, Compendium of Material Medica claims dendrobium candidum to have the effect of " the smart thick stomach of reinforcing yin essence benefit, never sufficient in benefit, longue meat, intelligence development, except frightened, made light of one's life by commiting suicide and prolonged life "; As a kind of rare medicinal herbs, it is listed in first of Chinese nine immortal grass in " Taoist Scriptures ".Confirm according to modern pharmacology experiment, the active ingredient mainly dendrobium polysaccharide of medicinal dendrobium, has significant curative effect to various diseases; At present, domestic existing tens of kinds take the stem of noble dendrobium as the Chinese patent drug product of raw material and compatibility, as Shiyu Yeguang Wan, stem of noble dendrobium sheet, JINSANGZI HOUPIAN, Mailuo Ning oral liquid, MAILUONING ZHUSHEYE, Yangyin Kouxiangye, dendrobium wine (Thousand Golden Prescriptions), the stem of noble dendrobium loose (" General Records of Holy Universal Relief ") etc., a lot of folk prescriptions of also useful stem of noble dendrobium disease therapy among the people.
Current stem of noble dendrobium market demand is very large; therefore wild Dendrodium is also excavated by a large amount of; endangered at present; in order to Conservation and development Dendrobium Plants; give full play to the resources advantage that the stem of noble dendrobium is huge; advance the development of stem of noble dendrobium industry, meet the demand of market to the stem of noble dendrobium, the plantation of the stem of noble dendrobium just seems of crucial importance.And the very suitable stem of noble dendrobium growth of the distinctive natural resources in Yunnan, low at Yunnan plantation stem of noble dendrobium cost, quality better, high efficiency, for this reason, Yunnan becomes domestic medicinal dendrobium planting scale and the maximum place of base quantity, and in the plantation of the stem of noble dendrobium, there is stem of noble dendrobium quality uneven, quality such as to can not be guaranteed at the problem, therefore, it is imperative for selecting pure, the applicable tame excellent dendrobium candidum new varieties of quality.
Because Seeds of Dendrobium Candidum is minimum, without endosperm, natural propagation rate is extremely low, and add artificial excessively collection and natural environment destruction, current China dendrobium officinale resource is endangered, is listed in Chinese Second Class Key Protected Plant.
In order to meet the needs of dendrobium candidum artificial planting to good seed, some stem of noble dendrobium kinds are by artificially breeding, by sexual propagation and vegetative mode, artificial planting is carried out to dendrobium candidum, as Zhejiang Province selects celestial No. 1, the dry measure used in former times of dendrobium candidum new varieties, celestial No. 2, dry measure used in former times, Guangdong Province selects No. 1, middle section, middle section is from all dendrobium candidums, Red River, Yunnan Province selects red prosperous No. 1, national institute of materia medica, Pu'er selects No. one, Pu'er, No. two etc., but these kinds are all some full sibs or family half sibs, namely after selfing or hybridization being carried out to specific parent, seedling colony of future generation is formed again with its capsule (including seed) breeding, as No. one, celestial dry measure used in former times, seed selection unit filters out and shows preferably Sxg-2, show obvious growth vigor, tissue-culturing rapid propagation has been carried out with the seed (capsule) of its artificial pollination, obtain a large amount of seedlings.The method of this sexual propagation because of dendrobium candidum be insect pollination under field conditions (factors), parental gene heterozygosis, between the offspring that selfing or hybridization obtain, genotype is heterogeneous, in the generation which results in same kind uniformity and from generation to generation between less stable.For vegetative propagation, bred, the trait segregation that the genetic recombination not occurring to cause due to both sexes Cell binding causes by the nutrition organs cuttage of same plant, press strip or tissue culture method, between individuality, proterties is relatively consistent, and from generation to generation, stability is higher.
Some stem of noble dendrobium kinds, by artificially breeding, being also used for the innovation of stem of noble dendrobium seed by Crossing system, as carried out artificial pollination when dendrobium candidum is bloomed, then selecting multiple parent's preparing hybrid combine, then are that explant expansion numerously obtains seedling by allocarpy implementation; Although the research utilizing the organ vegetative propagation such as stem apex, stem section to produce Multiple Buds acquisition regeneration plant has been reported, as domestic at the existing successfully report of the multiple stem of noble dendrobium such as iron sheet, Jin Chai, external Arditti etc. and Mosich etc. utilize stem of Dendrobium sections as explant, carry out stem of noble dendrobium vegetative propagation success, but cultivate stem of noble dendrobium Clonal Cultivar have not yet to see open report by combining both this.
Summary of the invention
The object of the present invention is to provide the breeding method of a kind of stem of noble dendrobium of the cultivation stem of noble dendrobium Clonal Cultivar adopting sexual propagation mode and vegetative manner to combine.
To achieve these goals, technical scheme of the present invention is:
A breeding method for the stem of noble dendrobium, completes according to step:
Step one, collects dendrobium candidum kind;
Step 2, selects general performance and carries out selfing as parent preferably, obtain capsule in collected dendrobium candidum kind;
Step 3, sows the capsule obtained on medium;
Step 4, to offspring's seedling seedling that medium bears, filters out the good individual plant of performance according to appearance index such as plant height, stem are thick, carries out asexual propagation to the individual plant of screening;
Step 5, obtains each clonal plant to step 4, carries out hardening, transplanting, obtains the clone stem of noble dendrobium.
In technique scheme, the medium of step 3 is: on the basis of 1/3MS medium, add kinetin KT that concentration is 0.1mg/L, 6-benzyl aminoadenine that concentration is 0.1mg/L and volume fraction be the murphy juice of 5%, wherein murphy juice adopts present known method to produce.
In technique scheme, in step 3, the incubation of capsule on medium is: temperature 23 ~ 28 DEG C, and intensity of illumination is 800 ~ 1200lux, and light application time is (8h ~ 10h)/24h, cultivates 90 days, within every 30 days, changes a subculture therebetween.Wherein, light application time refers to for (8h ~ 10h)/24h: illumination 8h ~ 10h in every 24h.
Preferably, in step 3, the incubation of capsule on medium is: temperature 25 DEG C, and intensity of illumination is 1000lux, and light application time is 8h/24h, cultivates 90 days, within every 30 days, changes a subculture therebetween.
In technique scheme, in step 4, asexual propagation is carried out to the stem of the individual plant of screening or root.
Particularly, adopt stem section to be the medium of explant asexual propagation protocorms to be: the basic element of cell division that concentration is the kinetin of 0.5mg/L, concentration is 1.5mg/L and mass fraction are the murphy juice of 10%, wherein, murphy juice adopts existing open method to prepare.
Particularly, medium when adopting root to carry out asexual propagation is: on the basis of 1/2MS medium, add methyl α-naphthyl acetate that concentration is 1mg/L, the basic element of cell division that indolebutyric acid that concentration is 1mg/L, concentration are 0.1mg/L and volume fraction be the bananas juice of 10%, wherein, bananas juice adopts existing open method to prepare.
The present invention is by hybridizing dendrobium candidum parent; After sowing capsule obtains the progeny population be separated, filter out fine individual plant according to appearance character and polyoses content; Carry out tissue cultures with its stem section or root as explant again and obtain seedling, merit is fixed, the stem of noble dendrobium Clonal Cultivar formed, be adapted at Yunnan to plant manyly, as the ground such as mountain of papers, Ban Na, Pu'er, the Baoshan, Dehong are planted, this, to the development promoting Yunnan stem of noble dendrobium industry, improves the situation of Yunnan Province's growing dendrobium kind confusion, enrich stem of noble dendrobium seed resource, the seed selection that promotion peasant household suits measures to local conditions has important value; Meanwhile, by the development of this stem of noble dendrobium kind plant husbandry, also can drive the development of regional economy, thus promote increasing peasant income, improve the living standard of peasant, be conducive to improving dendrobium candidum protection of resources, promote the development of China's Chinese Medicine Industry.
Embodiment
In order to understand the present invention better, illustrate content of the present invention further below in conjunction with embodiment, but content of the present invention is not only confined to the following examples.
The invention provides a kind of breeding method embodiment of the stem of noble dendrobium:
Step one, collect dendrobium candidum kind, the present invention collects Yunnan mountain of papers, Guangnan area count part dendrobium candidum germ plasm resource,
Step 2, selects general performance and carries out selfing as parent preferably, obtain capsule in collected dendrobium candidum kind; In embodiment, through cultivating collected dendrobium candidum kind, compare through between plant, select the good plant of general performance and carry out selfing as parent, wherein the good standard of general performance is: the appearance index such as plant height, stem stalk shape and sturdy degree, internode longitudinal grin color, leaf color, and filtering out the good dendrobium candidum SW3 of general performance as parent carries out selfing;
Step 3, cultivation is carried out in the capsule obtained sowing on medium and obtains seedling seedling, wherein, dendrobium candidum capsule selection standard is: choose October the ninety percent ripe uncracked capsule that kind of age is 150 days-160 days, described medium is on the basis of 1/3MS medium, add the kinetin KT that concentration is 0.1mg/L, concentration is the 6-benzyl aminoadenine of 0.1mg/L and volume fraction is the murphy juice of 5%, incubation is: by aseptic capsule planting seed on prior culture media, then temperature 25 DEG C, intensity of illumination is 1000lux, light application time is 8h/24h, cultivate 90 days, within every 30 days, change a subculture therebetween, obtain seedling seedling, wherein murphy juice described in medium adopts present known method to produce.
Step 4, to offspring's seedling seedling that medium bears, filters out performance good individual plant SW31, SW32, SW33, SW34, SW35, SW36, SW37, SW38 according to appearance index such as plant height, stem are thick, carries out asexual propagation to the individual plant of screening;
Wherein, asexual propagation can adopt stem section or rhizome, adopt stem section to be the medium that explant expands numerous protocorms to be: the basic element of cell division that concentration is the kinetin of 0.5mg/L, concentration is 1.5mg/L and mass fraction are the murphy juice of 10%, wherein, murphy juice adopts existing open method to prepare;
Medium when adopting root to carry out asexual propagation is: on the basis of 1/2MS medium, add methyl α-naphthyl acetate that concentration is 1mg/L, the basic element of cell division that indolebutyric acid that concentration is 1mg/L, concentration are 0.1mg/L and volume fraction be the bananas juice of 10%, wherein, bananas juice adopts existing open method to prepare;
Step 5, to each clonal plant that step 4 obtains, carries out hardening, transplanting, obtains the general performance good SW36 clone stem of noble dendrobium.
In embodiment, each clonal plant is obtained to step 4, transplant after in February, 2012, in March, 2012, in April, 2012 carrying out hardening respectively; And screen transplanting the stem of noble dendrobium grown up to from appearance character and polyoses content, select the good SW36 of general performance; And three batches of seedlings of SW36 are observed in stem stalk shape, internode longitudinal grin color, leaf color etc., its uniformity and stability are evaluated, this kind carries out polyoses content qualification through Institute of Analysis of Yunnan Province, data are in table five, meeting all standard of stem of noble dendrobium plant, is a choiceness strain.
Three batches of seedlings of SW36 are observed in stem stalk shape, internode longitudinal grin color, leaf color etc., the specificity of SW36, uniformity and stability are evaluated.
1. specificity
Table one be the kind SW36 that obtains of the present invention and parent SW3 about stem stalk color, internode longitudinal grin color be purple, spire front and the back side, mature leaf color contrast, kind SW36 the present invention obtained and parent SW3 cultivates under equivalent environment, adopt identical plantation specification and method, adopt the RHS plant colorimetric card contrast of British royal gardens association, SW36 stem stalk color is purple (PURPLE GROUP N97 B); Internode longitudinal grin color is purple (PURPLE GROUP N97 A); Spire front and the back side are all in purple (PURPLE GROUP N97 B); Mature leaf surface distribution purple plague purpura (PURPLE GROUP N97 A), limb edge is purple (PURPLE GROUP N97 A).
The kind SW36 that table one the present invention obtains and parent SW3 about stem stalk color, internode longitudinal grin color be purple, spire front and the back side, mature leaf color contrast.
Table one
2, uniformity and stability
In embodiment, each clonal plant is obtained to step 4, transplant after in February, 2012, in March, 2012, in April, 2012 carrying out hardening respectively, 100 Cong Miao that 3 crowdes are transplanted are observed, by there is good uniformity to aspects such as stem stalk color, young leaves color, mature leaf colors, between the generation, showed good stability, randomly drawed 30 clumps and observe from each colony, table two is the hardening stability datas of transplanting in February, 2012; Table three is the hardening stability datas of transplanting in March, 2012; Table four is the hardening stability datas of transplanting in April, 2012:
The hardening stability data of table two in February, 2012 transplanting
Table two
| Numbering | Stem stalk color | Young leaves color | Become leaf color |
| 1 | Purplish red | Purplish red | Spot is red |
| 2 | Purplish red | Purplish red | Spot is red |
| 3 | Purplish red | Purplish red | Spot is red |
| 4 | Purplish red | Purplish red | Spot is red |
| 5 | Purplish red | Purplish red | Spot is red |
| 6 | Purplish red | Purplish red | Spot is red |
| 7 | Purplish red | Purplish red | Spot is red |
| 8 | Purplish red | Purplish red | Spot is red |
| 9 | Purplish red | Purplish red | Spot is red |
| 10 | Purplish red | Purplish red | Spot is red |
| 11 | Purplish red | Purplish red | Spot is red |
| 12 | Purplish red | Purplish red | Spot is red |
| 13 | Purplish red | Purplish red | Spot is red |
| 14 | Purplish red | Purplish red | Spot is red |
| 15 | Purplish red | Purplish red | Spot is red |
| 16 | Purplish red | Purplish red | Spot is red |
| 17 | Purplish red | Purplish red | Spot is red |
| 18 | Purplish red | Purplish red | Spot is red |
| 19 | Purplish red | Purplish red | Spot is red |
| 20 | Purplish red | Purplish red | Spot is red |
| 21 | Purplish red | Purplish red | Spot is red |
| 22 | Purplish red | Purplish red | Spot is red |
| 23 | Purplish red | Purplish red | Spot is red |
| 24 | Purplish red | Purplish red | Spot is red |
| 25 | Purplish red | Purplish red | Spot is red |
| 26 | Purplish red | Purplish red | Spot is red |
| 27 | Purplish red | Purplish red | Spot is red |
| 28 | Purplish red | Purplish red | Spot is red |
| 29 | Purplish red | Purplish red | Spot is red |
| 30 | Purplish red | Purplish red | Spot is red |
The hardening stability data of table three in March, 2012 transplanting
Table three
| Numbering | Stem stalk color | Young leaves color | Become leaf color |
| 1 | Purplish red | Purplish red | Spot is red |
| 2 | Purplish red | Purplish red | Spot is red |
| 3 | Purplish red | Purplish red | Spot is red |
| 4 | Purplish red | Purplish red | Spot is red |
| 5 | Purplish red | Purplish red | Spot is red |
| 6 | Purplish red | Purplish red | Spot is red |
| 7 | Purplish red | Purplish red | Spot is red |
| 8 | Purplish red | Purplish red | Spot is red |
| 9 | Purplish red | Purplish red | Spot is red |
| 10 | Purplish red | Purplish red | Spot is red |
| 11 | Purplish red | Purplish red | Spot is red |
| 12 | Purplish red | Purplish red | Spot is red |
| 13 | Purplish red | Purplish red | Spot is red |
| 14 | Purplish red | Purplish red | Spot is red |
| 15 | Purplish red | Purplish red | Spot is red |
| 16 | Purplish red | Purplish red | Spot is red |
| 17 | Purplish red | Purplish red | Spot is red |
| 18 | Purplish red | Purplish red | Spot is red |
| 19 | Purplish red | Purplish red | Spot is red |
| 20 | Purplish red | Purplish red | Spot is red |
| 21 | Purplish red | Purplish red | Spot is red |
| 22 | Purplish red | Purplish red | Spot is red |
| 23 | Purplish red | Purplish red | Spot is red |
| 24 | Purplish red | Purplish red | Spot is red |
| 25 | Purplish red | Purplish red | Spot is red |
| 26 | Purplish red | Purplish red | Spot is red |
| 27 | Purplish red | Purplish red | Spot is red |
| 28 | Purplish red | Purplish red | Spot is red |
| 29 | Purplish red | Purplish red | Spot is red |
| 30 | Purplish red | Purplish red | Spot is red |
Table four is the hardening stability datas of transplanting in April, 2012
Table four
| Numbering | Stem stalk color | Young leaves color | Become leaf color |
| 1 | Purplish red | Purplish red | Spot is red |
| 2 | Purplish red | Purplish red | Spot is red |
| 3 | Purplish red | Purplish red | Spot is red |
| 4 | Purplish red | Purplish red | Spot is red |
| 5 | Purplish red | Purplish red | Spot is red |
| 6 | Purplish red | Purplish red | Spot is red |
| 7 | Purplish red | Purplish red | Spot is red |
| 8 | Purplish red | Purplish red | Spot is red |
| 9 | Purplish red | Purplish red | Spot is red |
| 10 | Purplish red | Purplish red | Spot is red |
| 11 | Purplish red | Purplish red | Spot is red |
| 12 | Purplish red | Purplish red | Spot is red |
| 13 | Purplish red | Purplish red | Spot is red |
| 14 | Purplish red | Purplish red | Spot is red |
| 15 | Purplish red | Purplish red | Spot is red |
| 16 | Purplish red | Purplish red | Spot is red |
| 17 | Purplish red | Purplish red | Spot is red |
| 18 | Purplish red | Purplish red | Spot is red |
| 19 | Purplish red | Purplish red | Spot is red |
| 20 | Purplish red | Purplish red | Spot is red |
| 21 | Purplish red | Purplish red | Spot is red |
| 22 | Purplish red | Purplish red | Spot is red |
| 23 | Purplish red | Purplish red | Spot is red |
| 24 | Purplish red | Purplish red | Spot is red |
| 25 | Purplish red | Purplish red | Spot is red |
| 26 | Purplish red | Purplish red | Spot is red |
| 27 | Purplish red | Purplish red | Spot is red |
| 28 | Purplish red | Purplish red | Spot is red |
| 29 | Purplish red | Purplish red | Spot is red |
| 30 | Purplish red | Purplish red | Spot is red |
Table five is stem of noble dendrobium plant polyose content appraising datums that Institute of Analysis of Yunnan Province cultivates the present invention.
Table five
| Test item | Unit | Testing result |
| Polysaccharide | g/100g | 61.8 |
| Moisture | g/100g | 84.2 |
The above; be only the specific embodiment of the present invention, but protection scope of the present invention is not limited thereto, is anyly familiar with those skilled in the art in the technical scope that the present invention discloses; change can be expected easily or replace, all should be encompassed within protection scope of the present invention.Therefore, protection scope of the present invention should described be as the criterion with the protection domain of claim.
Claims (2)
1. a breeding method for the stem of noble dendrobium, is characterized in that:
Complete according to following steps:
Step one, collects dendrobium candidum kind;
Step 2, filters out the good dendrobium candidum of general performance according to plant height, stem stalk shape and sturdy degree, internode longitudinal grin color and leaf color and carries out selfing as parent, obtain capsule in collected dendrobium candidum kind;
Step 3, by the capsule sowing that obtains on medium, described medium be on the basis of 1/3MS medium, add kinetin KT that concentration is 0.1mg/L, 6-benzyl aminoadenine that concentration is 0.1mg/L and volume fraction be the murphy juice of 5%; The incubation of described capsule on medium is: temperature 23 ~ 28 DEG C, and intensity of illumination is 800 ~ 1200lux, and light application time is 8h ~ 10h/24h, cultivates 90 days, within every 30 days, changes a subculture therebetween;
Step 4, to offspring's seedling seedling that medium bears, selects the good individual plant of performance according to plant height, stem scalping, carries out asexual propagation to the root of the individual plant of screening or stem section;
Adopt stem section to be the medium of explant asexual propagation protocorms to be: the basic element of cell division that concentration is the kinetin of 0.5mg/L, concentration is 1.5mg/L and mass fraction are the murphy juice of 10%;
Medium when adopting root to carry out asexual propagation is: on the basis of 1/2MS medium, add methyl α-naphthyl acetate that concentration is 1mg/L, the basic element of cell division that indolebutyric acid that concentration is 1mg/L, concentration are 0.1mg/L and volume fraction be the bananas juice of 10%;
Step 5, obtains each clonal plant to step 4, carries out hardening, transplanting, obtains the clone stem of noble dendrobium.
2. the breeding method of a kind of stem of noble dendrobium according to claim 1, it is characterized in that: in step 3, the incubation of capsule on medium is: temperature 25 DEG C, intensity of illumination is 1000lux, and light application time is 8h/24h, cultivate 90 days, within every 30 days, change a subculture therebetween.
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| CN101461328A (en) * | 2009-01-08 | 2009-06-24 | 中国科学院华南植物园 | High-efficient propagation method of seedlings of Dendrobium officinale |
| CN102283116A (en) * | 2011-06-29 | 2011-12-21 | 南开大学 | Simple industrial production method for dendrobium candidum |
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| JP2001299119A (en) * | 2000-04-25 | 2001-10-30 | Taku Sakurai | Method for creating dendrobium and dendrobium created by the same method |
| JP2010220596A (en) * | 2009-03-19 | 2010-10-07 | Nicchu Kokusai Tsusho Kk | Dendrobium growing downward |
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| CN101461328A (en) * | 2009-01-08 | 2009-06-24 | 中国科学院华南植物园 | High-efficient propagation method of seedlings of Dendrobium officinale |
| CN102283116A (en) * | 2011-06-29 | 2011-12-21 | 南开大学 | Simple industrial production method for dendrobium candidum |
| CN102388797A (en) * | 2011-07-21 | 2012-03-28 | 玉林市林业科学研究所 | Cross breeding method for Dendrobium officinale Kimura et Migo |
| CN102845308A (en) * | 2012-09-27 | 2013-01-02 | 浙江省中药研究所有限公司 | Method for dendrobium officinale tissue culture one-step seedling |
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