CN105010135A - Tissue culture and rapid propagation method of Polygonatum sibiricum - Google Patents
Tissue culture and rapid propagation method of Polygonatum sibiricum Download PDFInfo
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- CN105010135A CN105010135A CN201510367560.5A CN201510367560A CN105010135A CN 105010135 A CN105010135 A CN 105010135A CN 201510367560 A CN201510367560 A CN 201510367560A CN 105010135 A CN105010135 A CN 105010135A
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Abstract
The invention discloses a tissue culture and rapid propagation method of Polygonatum sibiricum. The method comprises the steps of explant selection and disinfection, inoculation culturing, propagation culturing, rooting culturing, and seedling hardening and transplanting. Concrete medium and culturing conditions in all the steps are screened, and a rapid propagation system of Polygonatum sibiricum is established, so the method is an effective approach for producing high-quality seedlings of Polygonatum sibiricum and popularizing cultivation.
Description
Technical field
The invention belongs to tissue culture rapid propagation technique field, be specifically related to a kind of tissue culture and rapid propagation method of sealwort.
Background technology
Sealwort is the dry rhizome of liliaceous plant sealwort (POlygonatumsibircum Red), David's-harp (P.Cyrtonema Hua) or P. kingianum (P.KingianumColl.etHemsl.).Different by medicinal material shape, practise and claim " polygonatum sibiricum Redoute ", " RHIZOMA POLYGONATI ZINGIBERIFORME ".Sealwort is put down, and taste is sweet.Effect of tool tonifying spleen moistening lung, supplementing qi and nourishing yin.Sealwort main product in Hebei, the Inner Mongol, the provinces and regions such as Shaanxi Province.David's-harp main product in Guizhou, Hunan, Yunnan, Anhui, the province such as Zhejiang.P. kingianum main product in Guizhou, Guangxi, the provinces and regions such as Yunnan.
Sealwort is medicine-food two-purpose medicinal material, wide market, in addition new drug development and health products exploitation, and the demand of people to sealwort also grows with each passing day, and wild resource can not meet need of production.A lot of place starts the artificial cultivation research that standardizes in recent years, but mainly carries out vegetative propagation with stem tuber, and reproduction coefficient is low, and the seminal propagation cycle is also longer, limits the production scale of sealwort.The rapid propagation system utilizing method for plant tissue culture to set up sealwort is the effective way of producing sealwort high quality seedling and cultivation popularization.
Summary of the invention
The object of the present invention is to provide a kind of tissue culture and rapid propagation method of sealwort, can accelerate the reproduction speed of sealwort, set up the rapid propagation system of sealwort, is the effective way of producing sealwort high quality seedling and cultivation popularization.
To achieve these goals, the invention provides following technical scheme:
A tissue culture and rapid propagation method for sealwort, comprises the steps:
(1) selection of explant and sterilization: choose and be with bud rhizome as explant, remove root system, peels off leaf bag sheet, cut terminal bud running water 1h, after terminal bud is dried on superclean bench with 75% alcohol-pickled 15 ~ 20s, after aseptic water washing 3 ~ 5 times, put into the HgCl of 0.2%
2in solution, take out after jog 5 ~ 8min, then use aseptic water washing 5 ~ 7 times;
(2) inoculated and cultured: be inoculated on inducing culture by the terminal bud of the explant through sterilization and cultivate, cultivates 30 ~ 35d and induces band bud tissue;
(3) Multiplying culture: by induction successfully band bud organize and be eachly cut into 5 ~ 8 parts, be then transferred to Multiplying culture proliferated culture medium carrying out 35 ~ 40d, breed differentiation simultaneously by the agglomerate of band indefinite bud and sprout;
(4) culture of rootage: until Multiplying culture break up the budlet seedling obtained grow to 2 ~ 4cm time, forward differentiation-inducing on root media taking root to, after 45d, grow 3 ~ 6 roots, root reaches 1 ~ 2cm;
(5) acclimatization and transplants: when the root of more than 90% bud seedling grow to more than 3, length >=1cm time, hardening can be carried out, be specially initial 1 ~ 2d blake bottle bottleneck first unlimited half that hardening starts, then bottleneck opens wide 2 ~ 3d entirely, select the seedling with more than 3 healthy and strong roots, carefully take out with tweezers, then wash agar off with clear water, seedling is transplanted in the seedling medium of seedling-cultivating tray in hardening booth, cover light pressure with soil, plant well wetting with sprayer water spray, finally tame, ventilate, management of sheltering from heat or light, the hardening time is 20-30d, can transplant.
In the tissue culture and rapid propagation method of aforementioned sealwort, before carrying out step (4) described culture of rootage, change bottle and repeat step (3) described Multiplying culture, to obtain more polygerm block.
In the tissue culture and rapid propagation method of aforementioned sealwort, the inducing culture described in step (2) is MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 5.8 ~ 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 1800 ~ 2200lx, and light application time is 14h/d; Preferably, the pH value of described inducing culture is 6.0; Intensity of illumination in condition of culture is 2000lx.
In the tissue culture and rapid propagation method of aforementioned sealwort, the proliferated culture medium described in step (3) is MS+6-BA2.0mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, and light application time is 14h/d.
In the tissue culture and rapid propagation method of aforementioned sealwort, the root media described in step (4) is MS+NAA1.0mg/l+IBA0.5mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, and light application time is 14h/d.
In the tissue culture and rapid propagation method of aforementioned sealwort, the described hardening canopy temperature of step (5) controls at 20 ~ 30 DEG C, and relative moisture controls between 75% ~ 85%.
In the tissue culture and rapid propagation method of aforementioned sealwort, the seedling medium described in step (5) is: river sand, ash and peat soil carry out the mixture mixed according to the volume ratio of 1:1:1.
In order to ensure beneficial effect of the present invention, applicant carried out a series of experiment, specific as follows:
One, the selection of explant
Select the terminal bud of sealwort and the tender stem tuber of children of Xin Fa to cultivate as explant, condition of culture is unanimously temperature 26 DEG C, and intensity of illumination is 2000lx, and light application time is 14h/d; Medium is MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 5.8; Appear for the first time with regard to its rate of increase and indefinite bud and contrast day, the results are shown in Table 1.From comparing result, select terminal bud as explant, with the tender stem tuber of children as compared with explant, its rate of increase is higher, and indefinite bud appears day for the first time comparatively early.
Table 1 different explants contrasts
| Explant | The rate of increase (%) | Indefinite bud appears day (d) for the first time |
| Terminal bud | 11.7 | 20.8d |
| The tender stem tuber of children | 8.1 | 22.1d |
Two, HgCl
2the selection of concentration
By HgCl
2bacteria-removing liquid three kinds of concentration 0.1%, 0.2%, 0.3% and different degerming time contrast, and best when we find selection 0.2% concentration 5min, living contaminants is minimum, and excessive concentration and overlong time also can kill sealwort tissue simultaneously, are lowered into motility rate.
Three, the selection of hormone kind and concentration
(1) inducing culture: take proliferative induction to differentiate indefinite bud, devises during cultivation in MS medium and adds sucrose 25g/l and agar 5g/l, then carries out comparative trial with regard to hormon and proportioning thereof, is specifically respectively:
A1，NAA0.1+KT1.0；
A2，NAA0.2+KT1.0；
A3，NAA0.3+KT1.0；
A4，6-BA2.0+NAA0.2+KT1.0；
A5，6-BA2.5+NAA0.2+KT1.0；
A6，6-BA2.5+NAA0.2+KT2.0；
A7，6-BA2.5+NAA0.1；
A8，6-BA2.5+NAA0.2；
A9，6-BA2.5+NAA0.3。
Above unit is mg/l, and find by testing us, KT and 6-BA is obvious for the proliferation function of tissue, A2 indefinite bud appears day for the first time, and early still differentiation rate is relative with proliferation times low, A5 and A6 differentiation and proliferation is higher, but A6 differentiation is lower than A5, wherein A5 differentiation and proliferation is best, and the differentiation rate of bud reaches 10.8.Illustrate that relative concentration is the quick differentiation that the 6-BA of 2.5mg/l is conducive to hyperblastosis and bud.
(2) proliferated culture medium: the Proliferation, Differentiation of clump bud is cultivated and selected MS medium to add sucrose 25g/l and agar 5g/l, carries out comparative trial, be specially hormon concentration:
B1，6-BA1.0+NAA0.2+KT1.0；
B2，6-BA2.0+NAA0.2+KT1.0；
B3，6-BA3.0+NAA0.2+KT1.0；
B4，6-BA2.0+NAA0.2+KT2.0；
B5，6-BA2.5+NAA0.2+KT2.5；
B6，6-BA3.0+NAA0.2+KT2.0；
We find, more high proliferation multiple is higher for 6-BA concentration, but when reaching 3.0mg/l, proliferation times reduces on the contrary, and the generating period of B2 and B6 is relatively short, is respectively 35.8d and 35.6d, and with B2 proliferation times for 5.58, generating period 35.8d is best.
(3) root media: MS medium adds sucrose 25g/l and agar 5g/l, then adds (0.1,0.3 respectively, 0.5,0.8) mg/l IBA and (0.6,0.8,1.0,1.2) mg/l NAA contrasts, found by test, the concentration of IBA and NAA has remarkable impact to the incidence of its root and root number within the specific limits, best with 0.5mg/L IBA and 1.0mg/LNAA respectively, rooting rate reaches 95%, and both are all not remarkable on the impact of root long number.
Four, the selection of seedling medium
We choose different substrates proportioning and contrast to transplant hardening, specific as follows:
D1: peat soil: river sand=2:1;
D2: ash: river sand=2:1;
D3: peat soil: ash: river sand=1:1:1;
Seedling is transplanted in above-mentioned three kinds of matrix, canopy temperature controls at 20 ~ 30 DEG C, relative moisture is between 75% ~ 85%, after 30d, we find that the survival rate of D2 group is relatively low, be 70.5%, the survival rate of D1 and D3 all reaches more than 91%, and new root incidence is respectively 84% and 95%, so preferably D3 is seedling medium.
The invention has the beneficial effects as follows:
The present invention utilizes method for plant tissue culture to establish the rapid propagation system of sealwort, is the effective way of producing sealwort high quality seedling and cultivation popularization.
The present invention has carried out experiment sieving and optimization to various medium, consumption and the condition of culture used in sealwort reproductive process, effectively raises the reproduction coefficient of sealwort.
Incubation speed of the present invention is fast, and regeneration frequency is higher, and aberration rate is lower, and the cycle is relatively short, can keep the genetic stability of sealwort preferably.
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
Embodiment 1
A tissue culture and rapid propagation method for sealwort, comprises the steps:
(1) selection of explant and sterilization: choose and be with bud rhizome as explant, remove root system, peels off leaf bag sheet, cut terminal bud running water 1h, after terminal bud is dried on superclean bench with 75% alcohol-pickled 15s, after aseptic water washing 3 times, put into the HgCl of 0.2%
2in solution, take out after jog 5min, then use aseptic water washing 5 times;
(2) inoculated and cultured: the terminal bud of the explant through sterilization is inoculated into inducing culture (MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, pH value is adjusted to 5.8) on cultivate, condition of culture is: temperature 26 DEG C, intensity of illumination 1800lx, light application time is 14h/d; Cultivate about 30d and induce band bud tissue;
(3) Multiplying culture: by induction successfully band bud organize and be eachly cut into 5 parts, then Multiplying culture proliferated culture medium (MS+6-BA2.0mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, pH value is adjusted to 6.0) carrying out 35d is transferred to; Condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, and light application time is 14h/d, breeds differentiation simultaneously sprout by the agglomerate of band indefinite bud;
(4) culture of rootage: until Multiplying culture break up the budlet seedling obtained grow to 2 ~ 4cm time, forward root media (MS+NAA1.0mg/l+IBA0.5mg/l+ sucrose 25g/l+ agar 5g/l to, pH value is adjusted to 6.0) go up differentiation-inducing taking root, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d, and after 45d, grow 3 ~ 6 roots, root reaches about 1cm;
(5) acclimatization and transplants:
Matrix prepares: raise seedling in greenhouse matrix river sand, ash and peat soil mix according to the volume ratio of 1:1:1, put into seedling-cultivating tray and strike off, and uses water saturates matrix before using;
When the root of more than 90% bud seedling grow to more than 3, length >=1cm time, hardening can be carried out, be specially the initial 1d blake bottle bottleneck first unlimited half that hardening starts, then bottleneck opens wide 3d entirely, select the seedling with more than 3 healthy and strong roots, carefully take out with tweezers, then wash agar off with clear water, seedling is transplanted in the seedling medium of seedling-cultivating tray in hardening booth, cover light pressure with soil, plant well wetting with sprayer water spray, finally tame, ventilate, carry out with shade net management of sheltering from heat or light, the hardening time is 20d, can transplant.
As a result, carry out the Fast-propagation of sealwort as stated above, the increment multiple in each cycle reaches 64.6, and the root induction rate in culture of rootage reaches 96%, the survival rate 93% of seedling after acclimatization and transplants of taking root.
Embodiment 2
A tissue culture and rapid propagation method for sealwort, comprises the steps:
(1) selection of explant and sterilization: choose and be with bud rhizome as explant, remove root system, peels off leaf bag sheet, cut terminal bud running water 1h, after terminal bud is dried on superclean bench with 75% alcohol-pickled 18s, after aseptic water washing 4 times, put into the HgCl of 0.2%
2in solution, take out after jog 6min, then use aseptic water washing 6 times;
(2) inoculated and cultured: the terminal bud of the explant through sterilization is inoculated into inducing culture (MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, pH value is adjusted to 6.0) on cultivate, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d; Cultivate about 33d and induce band bud tissue;
(3) Multiplying culture: by induction successfully band bud organize and be eachly cut into 6 parts, then proliferated culture medium (MS+6-BA2.0mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l is transferred to, pH value is adjusted to 6.0) on carry out the Multiplying culture of 38d, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d, breeds differentiation simultaneously sprout by the agglomerate of band indefinite bud; Then change bottle and repeat this step Multiplying culture, to obtain more polygerm block;
(4) culture of rootage: until Multiplying culture break up the budlet seedling obtained grow to 2 ~ 4cm time, forward root media (MS+NAA1.0mg/l+IBA0.5mg/l+ sucrose 25g/l+ agar 5g/l to, pH value is adjusted to 6.0) go up differentiation-inducing taking root, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d, and after 45d, grow 3 ~ 6 roots, root reaches 1 ~ 2cm;
(5) acclimatization and transplants:
Matrix prepares: raise seedling in greenhouse matrix river sand, ash and peat soil mix according to the volume ratio of 1:1:1, put into seedling-cultivating tray and strike off, and uses water saturates matrix before using;
When the root of more than 90% bud seedling grows to more than 3, during length >=1cm, hardening can be carried out, be specially the initial 2d blake bottle bottleneck first unlimited half that hardening starts, then bottleneck opens wide 3d entirely, select the seedling with more than 3 healthy and strong roots, carefully take out with tweezers, then agar is washed off with clear water, (temperature of shed controls at 20 ~ 30 DEG C seedling to be transplanted to hardening booth, relative moisture controls between 75% ~ 85%) in seedling-cultivating tray seedling medium in, light pressure is covered with soil, plant well wetting with sprayer water spray, finally tame, ventilate, management of sheltering from heat or light is carried out with shade net, the hardening time is 25d, can transplant.
As a result, carry out the Fast-propagation of sealwort as stated above, the proliferation times in each cycle reaches 63.6, and the root induction rate in culture of rootage reaches 95%, the survival rate 92% of seedling after acclimatization and transplants of taking root.
Embodiment 3
A tissue culture and rapid propagation method for sealwort, comprises the steps:
(1) selection of explant and sterilization: choose and be with bud rhizome as explant, remove root system, peels off leaf bag sheet, cut terminal bud running water 1h, after terminal bud is dried on superclean bench with 75% alcohol-pickled 20s, after aseptic water washing 5 times, put into the HgCl of 0.2%
2in solution, take out after jog 8min, then use aseptic water washing 7 times;
(2) inoculated and cultured: the terminal bud of the explant through sterilization is inoculated into inducing culture (MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, pH value is adjusted to 5.8) on cultivate, condition of culture is: temperature 26 DEG C, intensity of illumination 2200lx, light application time is 14h/d; Cultivate about 35d and induce band bud tissue;
(3) Multiplying culture: by induction successfully band bud organize and be eachly cut into 8 parts, then proliferated culture medium (MS+6-BA2.0mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l is transferred to, pH value is adjusted to 6.0) on carry out the Multiplying culture of 40d, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d, breeds differentiation simultaneously sprout by the agglomerate of band indefinite bud; Then change bottle and repeat this step Multiplying culture, to obtain more polygerm block;
(4) culture of rootage: until Multiplying culture break up the budlet seedling obtained grow to 2 ~ 4cm time, forward root media (MS+NAA1.0mg/l+IBA0.5mg/l+ sucrose 25g/l+ agar 5g/l to, pH value is adjusted to 6.0) go up differentiation-inducing taking root, condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, light application time is 14h/d, and after 45d, grow 3 ~ 6 roots, root reaches about 2cm;
(5) acclimatization and transplants:
Matrix prepares: raise seedling in greenhouse matrix is put into seedling-cultivating tray and strikes off, and uses water saturates matrix before using;
When the root of more than 90% bud seedling grows to more than 3, during length >=1cm, hardening can be carried out, be specially the initial 2d blake bottle bottleneck first unlimited half that hardening starts, then bottleneck opens wide 2d entirely, select the seedling with more than 3 healthy and strong roots, carefully take out with tweezers, then agar is washed off with clear water, (temperature of shed controls at 20 ~ 30 DEG C seedling to be transplanted to hardening booth, relative moisture controls between 75% ~ 85%) in seedling-cultivating tray seedling medium in, light pressure is covered with soil, plant well wetting with sprayer water spray, finally tame, ventilate, to shelter from heat or light management, the hardening time is 30d, can transplant.
As a result, carry out the Fast-propagation of sealwort as stated above, the proliferation times in each cycle reaches 62.5, and the root induction rate in culture of rootage reaches 96%, the survival rate 94% of seedling after acclimatization and transplants of taking root.
Claims (8)
1. a tissue culture and rapid propagation method for sealwort, is characterized in that, comprises the steps:
(1) selection of explant and sterilization: choose and be with bud rhizome as explant, remove root system, peels off leaf bag sheet, cut terminal bud running water 1h, after terminal bud is dried on superclean bench with 75% alcohol-pickled 15 ~ 20s, after aseptic water washing 3 ~ 5 times, put into the HgCl of 0.2%
2in solution, take out after jog 5 ~ 8min, then use aseptic water washing 5 ~ 7 times;
(2) inoculated and cultured: be inoculated on inducing culture by the terminal bud of the explant through sterilization and cultivate, cultivates 30 ~ 35d and induces band bud tissue;
(3) Multiplying culture: by induction successfully band bud organize and be eachly cut into 5 ~ 8 parts, be then transferred to Multiplying culture proliferated culture medium carrying out 35 ~ 40d, breed differentiation simultaneously by the agglomerate of band indefinite bud and sprout;
(4) culture of rootage: until Multiplying culture break up the budlet seedling obtained grow to 2 ~ 4cm time, forward differentiation-inducing on root media taking root to, after 45d, grow 3 ~ 6 roots, root reaches 1 ~ 2cm;
(5) acclimatization and transplants: when the root of more than 90% bud seedling grow to more than 3, length >=1cm time, hardening can be carried out, be specially initial 1 ~ 2d blake bottle bottleneck first unlimited half that hardening starts, then bottleneck opens wide 2 ~ 3d entirely, select the seedling with more than 3 healthy and strong roots, carefully take out with tweezers, then wash agar off with clear water, seedling is transplanted in the seedling medium of seedling-cultivating tray in hardening booth, cover light pressure with soil, plant well wetting with sprayer water spray, finally tame, ventilate, management of sheltering from heat or light, the hardening time is 20-30d, can transplant.
2. the tissue culture and rapid propagation method of sealwort according to claim 1, is characterized in that, before carrying out step (4) described culture of rootage, changes bottle and repeats step (3) described Multiplying culture, to obtain more polygerm block.
3. the tissue culture and rapid propagation method of sealwort according to claim 1 and 2, is characterized in that, the inducing culture described in step (2) is MS+6-BA2.5mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 5.8 ~ 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 1800 ~ 2200lx, and light application time is 14h/d.
4. the tissue culture and rapid propagation method of sealwort according to claim 3, is characterized in that, the pH value of described inducing culture is 6.0; Intensity of illumination in condition of culture is 2000lx.
5. the tissue culture and rapid propagation method of sealwort according to claim 1 and 2, is characterized in that, the proliferated culture medium described in step (3) is MS+6-BA2.0mg/l+NAA0.2mg/l+KT1.0mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, and light application time is 14h/d.
6. the tissue culture and rapid propagation method of sealwort according to claim 1 and 2, is characterized in that, the root media described in step (4) is MS+NAA1.0mg/l+IBA0.5mg/l+ sucrose 25g/l+ agar 5g/l, and its pH value is 6.0; Condition of culture is: temperature 26 DEG C, intensity of illumination 2000lx, and light application time is 14h/d.
7. the tissue culture and rapid propagation method of sealwort according to claim 1 and 2, is characterized in that, the described hardening canopy temperature of step (5) controls at 20 ~ 30 DEG C, and relative moisture controls between 75% ~ 85%.
8. the tissue culture and rapid propagation method of sealwort according to claim 1 and 2, is characterized in that, the seedling medium described in step (5) is: river sand, ash and peat soil carry out the mixture mixed according to the volume ratio of 1:1:1.
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| CN113100057A (en) * | 2021-04-02 | 2021-07-13 | 安徽省林业高科技开发中心 | Seedling strengthening and rapid propagation method for polygonatum cyrtonema |
| CN115005038A (en) * | 2022-07-16 | 2022-09-06 | 江西农业大学 | Method for rapidly inducing multiple buds of rhizoma polygonati |
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| CN105580734A (en) * | 2016-01-14 | 2016-05-18 | 遵义市龙驰生物科技有限公司 | Efficient inducing method of in-vitro rhizomes of Polygonatum sibiricum Red |
| CN109302983A (en) * | 2017-07-26 | 2019-02-05 | 湖南省林业科学院 | It is a kind of using terminal bud as the polygonatum cyrtonema rapid propagation method of explant |
| CN107466768A (en) * | 2017-08-22 | 2017-12-15 | 湖口县顺昌中药材有限公司 | The method that sealwort nursery carries with cape jasmine set |
| CN110463590B (en) * | 2018-12-27 | 2021-04-06 | 广西壮族自治区农业科学院生物技术研究所 | Method for promoting germination of polygonatum cyrtonema rooted seedlings |
| CN110463590A (en) * | 2018-12-27 | 2019-11-19 | 广西壮族自治区农业科学院生物技术研究所 | A kind of method that polygonatum cyrtonema rooted seedling promotees bud |
| CN110741937A (en) * | 2019-11-27 | 2020-02-04 | 绍兴文理学院 | Rapid propagation method of polygonatum sibiricum |
| CN110741937B (en) * | 2019-11-27 | 2021-02-19 | 绍兴文理学院 | Rapid propagation method of polygonatum sibiricum |
| CN112042521A (en) * | 2020-08-10 | 2020-12-08 | 阳山县三连阳生态农林开发有限公司 | Transplanting method for test-tube plantlet of rhizome plant |
| CN113100057A (en) * | 2021-04-02 | 2021-07-13 | 安徽省林业高科技开发中心 | Seedling strengthening and rapid propagation method for polygonatum cyrtonema |
| CN115005038A (en) * | 2022-07-16 | 2022-09-06 | 江西农业大学 | Method for rapidly inducing multiple buds of rhizoma polygonati |
| CN116530415A (en) * | 2023-05-30 | 2023-08-04 | 贵州省植物园 | Circularly operated polygonatum cyrtonema tissue culture breeding method |
| CN116530415B (en) * | 2023-05-30 | 2024-04-02 | 贵州省植物园 | Circularly operated polygonatum cyrtonema tissue culture breeding method |
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