CN103404442A - Induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther - Google Patents
Induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther Download PDFInfo
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- CN103404442A CN103404442A CN2013103552663A CN201310355266A CN103404442A CN 103404442 A CN103404442 A CN 103404442A CN 2013103552663 A CN2013103552663 A CN 2013103552663A CN 201310355266 A CN201310355266 A CN 201310355266A CN 103404442 A CN103404442 A CN 103404442A
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Abstract
The invention discloses an induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther, which belongs to the field of plant tissue culture techniques. The method comprises the following steps: (1) preparation of a medium; (2) somatic embryogenesis; (3) germination and seedling formation of a somatic embryo; and (4) domestication, transplantation and the like of a tissue culture seedling. According to the invention, the induction method is a tissue culture method established with the macleaya cordata anther as an explant and can realize rapid, high-efficiency and large-scale production of high-quality macleaya cordata haploid plants; after field screening, the macleaya cordata haploid plants can be directly used as breeding materials for research on breeding and genomics.
Description
Technical field
The present invention relates to field of plant tissue culture technique, be specifically related to a kind of method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration.
Background technology
Macleaya cordata (Macleaya cordata (Willd) R.Br) is Papaveraceae Macleaya plant, be rich in multiple isoquinoline alkaloid, as sanguinarine (sanguinarine, SA), Chelerythrine (chelerythrine, CHE), Biflorine (protopine, PRO) and allocryptopine (allocryptopine, ALL) etc. (Guo Yuge, once founded the state, Tan Manliang, Deng. in macleaya cordata leaf and macleaya microcarpa Fedde, 4 kinds of alkaloidal content are relatively. Central-South pharmacy .2011,9(11): 829-832).They have significantly and abundant biologically active and pharmacologically active; at the gut flora (Cai Peng that regulates pig; Sun Zhiliang; once founded the state; Deng. the impact of various dose macleaya cordata extracts on Growth Performance of Weaning Piglets. Chinese animal and veterinary .2010,37(5): 41-43), improve liver function (Xiao Li, Yi Jian; Zhao Jing, etc. the protective effect of macleaya cordata extracts to acute liver injury of rats. Central-South pharmacy.2011,9(7): 486-490), sterilization (institute of Chinese materia medica, Shanghai. the bacteriostatic test of macleaya cordata and animal experiment [R] .1977), antitumor (Fan Shulian, appoint peak, Zhang Yuan, Deng. effect research [J] Shaanxi tumour medical science of macleaya cordata total alkaloid to animal transplanting tumor, 174) etc. 2000,8(3): aspect has broad application prospects.Simultaneously, the biomass of macleaya cordata is larger, and fuel value is higher, is a kind of important energy-source plant, and Germany scientist been has has been researched and developed and gone into operation based on the energy products of macleaya cordata Biomass Energy Technology.At present, increasing by the product that macleaya cordata is developed, feed addictive, bactericide, mouthwash, botanical pesticide etc. are arranged; 2011, the loose country's " new veterinary drug certificate " (two classes) that obtains respectively of the macleaya cordata extracts of Hunan Micolta Bioresource Inc.'s development and macleaya cordata, filled up domestic blank in two class veterinary drug research and development especially.Along with the demand of market to macleaya cordata extracts and macleaya cordata itself increases day by day, supply falls short of demand just to have caused the macleaya cordata raw material.How to improve particularly alkaloidal output in macleaya cordata of macleaya cordata, become a problem demanding prompt solution, more become the bottleneck of restriction macleaya cordata industry development.Along with the price of labour power, rent of soil constantly raises, and the integrated cost that will inevitably cause researching and developing macleaya cordata by the cultivated area that increases macleaya cordata merely increases greatly, and this has just restricted the input of macleaya cordata raw material to market greatly.Therefore must on the basis of implant mass, carry out the biological engineering to obtaining the higher kind of alkaloid to macleaya cordata.The haplobiont obtained by the macleaya cordata vitro anther culture is the important colony basis that macleaya cordata is carried out breeding improvement.The haplobiont that adopts the macleaya cordata vitro anther culture to obtain is carried out breeding can shortening the breeding cycle, overcomes the incompatibility of distant hybridization, improves the efficiency of mutation breeding, the accuracy that also can greatly improve gene location, mark on a map simultaneously.At present, anther culture has not only become studying physiological, biochemical and important method that heredity is commonly used, and for the very important meaning of having illustrated of gene regulation, expression mechanisms in experimental embryology, physiology and molecular biology.
The vitro anther culture of plant starts from 20 century 70s, along with the continuous maturation of technology and going deep into gradually of research, have at present on many plants and by vitro anther culture, successfully obtained monoploid, wood (Li Yanlin continues if red, Yu Xiaoying, Xiong Xingyao, Deng continue inducing of wooden pollen monoploid callus and cultivate [J] of. safflower. Agricultural University Of Hunan's journal (natural science edition), 2011, 37(6): 633), apple (Xue Guangrong. the pollen plant of 8 main breeds of apple anther culture technique is cultivated successfully [J]. Scientia Agricultura Sinica, 1990(3): 86), oranges and tangerines (Germanna M A, Reforgiato G.Hploid embryos regeneration from another culture of ' Mapo ' tangelo[J] .Advan Hort sci, 1997 (11): 147-152), Lee (Sun Jianyun, Wang Qingya. the growth of Lee's flower pesticide pollen and Histochemical studies [J]. the Agriculture of Anhui science, 2005, 33 (8): 1402-1404), lichee (Fu Lianfang. the research of lichee pollen plant induction. Acta Genetica Sinica, 1983, 10(5): 369-374), watermelon (Yuan Wanliang. watermelon flower training Preliminary Report on Experiment. the Shaanxi agricultural science, 1995(1): 29-30), loquat (Maria, Antonietta, Germana.Development of multicellular pollen of Eriobotrya japonica Lindl.through another culture[J], PLANT SCIENCE, 2006 (171): 718-725), asparagus (Peng Xinhong. the pre-test [J] of asparagus anther culture embryoid induction condition optimizing. Agricultural University Of Jiangxi's journal, 2006, 28(1) 39-43), tomato (high high official position. the tomato vitro anther obtains plant [J]. the gardening journal, 1980, 7 (4): 37-41) and capsicum (Zhang Enrang, Xu Haili. the Primary Study (J) of Guizhou Native pepper anther culture system. the capsicum magazine, 2005(4): 24-27) etc.But also do not find up to now about macleaya cordata vitro anther culture success Haploid production plant report.
Summary of the invention
The objective of the invention is, for market, adopt seminal propagation the differentiation of descendant inheritting proterties and the contradiction that modes of reproduction can not be met the need of market to occur to good macleaya cordata seed resource, a kind of method of utilizing tissue culture technique production macleaya cordata monoploid seedling is provided, overcome the existing existing defect of propagation method, be convenient to the high-quality macleaya cordata germ plasm resource of quick, efficient, large-scale seed selection heredity homogeneous, health, be applied to macleaya cordata breeding and genomics research.
Purpose of the present invention realizes by following technical scheme.
A kind of method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration, comprise the steps:
1) the taking and sterilizing of explant: get and be in full-bloom stage, length, after the macleaya cordata bud sterilization of 2-12mm, pushes bud aside gently, the stamen of choosing, and the excision filigree, stay flower pesticide as explant, standby;
2) embryo callus induce cultivation: the flower pesticide of step 1) sterilization is inoculated into to BLCG+2,4-D0.5 on～3.0mg/L+6-BA0.5mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000～3000Lx, under the condition of light application time 16～18h/d, carry out inducing of embryo callus, obtain callus after cultivation 30～40d;
3) after callus switching flower pesticide embryoid induction cultivation: by step 2) obtained, form embryoid;
4) somatic embryo is sprouted with seedling and is cultivated: form the whole plant of taking root after the embryoid switching that step 3) is obtained;
5) transplanting of regrowth and cultivation: will take root transplantation of seedlings after hardening in hardening matrix, then carry out normal water and fertilizer management.
The process of taking with sterilizing of the described explant of step 1) is specially: get and be in full-bloom stage, length at the macleaya cordata bud of 2-12mm, is cleaned with distilled water, clean with aseptic water washing on superclean bench; With 70% alcohol-pickled 10s, then use aseptic water washing 3～4 times after with 0.1% mercuric chloride solution, soaking 8～10min sterilization; Bud is pushed aside gently, the stamen of choosing, the excision filigree, stay flower pesticide as explant, is put in suck dry moisture on the filter paper of aseptic drying, standby.
The described flower pesticide embryoid induction of step 3) cultivation process is specially: by step 2) callus that obtains is transferred to MS+2,4-D0.5 on～1mg/L+6-BA0.5mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, under the condition of light application time 16-18h/d, carry out the cultivation of inducing of embryoid, form embryoid after 20 days.
The described somatic embryo of step 4) is sprouted with the seedling cultivation process and is: the embryoid that step 3) is obtained is transferred to MS+GA
3On 0～1mg/L+IAA0～0.25mg/L+KT0～0.25mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, cultivate under the condition of light application time 16～18h/d and formed the whole plant of taking root in 10～15 days.
The detailed process of step 5) is as follows: the seedling of taking root, be transplanted to peat soil: wood peat soil: in the hardening matrix of coconut palm chaff powder: vermiculite=1:1:1:1, water permeable, and cover seedling with the plastics hut, guarantee that canopy class humidity is not less than 80%, 25 ± 1 ℃ of temperature, intensity of illumination is to cultivate under 3000lx, seedling adapts to culture environment after two weeks, removes plastic greenhouse, carries out normal water and fertilizer management.
Preferred in said method:
Embryonic callus induction medium: BLCG+2,4-D0.5mg/L+6-BA0.5mg/L;
Flower pesticide embryoid induction medium: MS+2,4-D0.5mg/L+6-BA0.5mg/L;
Somatic embryo is sprouted and seedling medium: MS+GA
30.5mg/L.
This invention has the following advantages:
(1) using flower pesticide as explant material, pollution rate is low, and the callus of induce rate is high;
(2) the macleaya cordata anther tissue somatic embryo generation tissue culture technique of setting up, can overcome the differentiation of current seminal propagation characters of progenies, genetic character is unstable with root, splits reproduction coefficient defect on the low side, can cultivate in the short period of time that genotype is isozygotied, the macleaya cordata germ plasm resource of inheritance stability, can apply widely the traditional breeding method of macleaya cordata and the research of biotechnology.
The accompanying drawing explanation
Fig. 1 is that in the macleaya cordata anther tissue, sediments microscope inspection is in the keep to the side photo of phase microspore cell of monokaryon;
Fig. 2 is that flower pesticide induces cultivation to produce the photo that embryo callus lures;
Fig. 3 is that macleaya cordata flower pesticide produces the photo of embryoid by the microspore embryo callus;
The embryoid of Fig. 4 for sprouting;
The embryoid of Fig. 5 for having sprouted;
Fig. 6 is the macleaya cordata regrowth that rooting culture survives.
Embodiment
The present invention is described in further detail by following examples, but content of the present invention is not limited to this.
Embodiment 1
(1) preparation of medium comprises component and proportioning thereof that minimal medium and tissue are cultivated each stage medium:
1) minimal medium: minimal medium has two kinds of BLCG and MS, wherein adds respectively sucrose 30g/L, agar 7g/L, pH5.8;
2) embryo callus medium: BLCG+2,4-D0.5mg/L+6-BA0.5mg/L;
3) embryoid induction medium: MS+2,4-D0.5mg/L+6-BA0.5mg/L;
4) somatic embryo is sprouted and seedling medium: MS+GA
30.5mg/L;
(2) the anther tissue somatic embryo occurs:
1) the taking and sterilizing of explant: get and be in full-bloom stage, length is at 2-12mm, (the most of microspore cells of microscopic examination are in the keep to the side flower pesticide part of phase of monokaryon to the macleaya cordata bud that color is emerald green, referring to accompanying drawing 1), with distilled water, clean, clean with aseptic water washing on superclean bench; With 70% alcohol-pickled 10s, then use aseptic water washing 3～4 times after with 0.1% mercuric chloride solution, soaking 8～10min sterilization; Bud is pushed aside gently, the stamen of choosing, the excision filigree, stay flower pesticide as explant, is put in suck dry moisture on the filter paper of aseptic drying, standby;
2) embryo callus induce cultivation: above-mentioned flower pesticide is inoculated into to step (1) 2) on the embryonic callus induction medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, under the condition of light application time 16h/d, carry out inducing of embryo callus, after cultivating 30～40d, obtain well-grown callus (referring to accompanying drawing 2), its callus of induce rate can reach 16.79%, and growth coefficient reaches as high as 3;
3) flower pesticide embryoid induction cultivation: above-mentioned callus is transferred to (1) 3) on the inducing culture of flower pesticide embryoid, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, under the condition of light application time 16h/d, carry out the cultivation of inducing of embryoid, after 20 days, form embryoid (referring to accompanying drawing 3), the embryoid induction rate reaches as high as 19.65%, and growth coefficient reaches as high as 4;
(3) somatic embryo is sprouted and seedling is cultivated: above-mentioned embryoid is transferred to step (1) 4) somatic embryo sprout with the seedling medium on, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, cultivate 10～15 days (referring to accompanying drawing 4,5) under the condition of light application time 16h/d;
(4) domestication of group training seedling and transplanting: the seedling of taking root shifts out with tweezers, be transplanted to peat soil: wood peat soil: coconut palm chaff powder: in vermiculite=1:1:1:1 hardening matrix, water permeable, and cover seedling with the plastics hut, guarantee that canopy class humidity is not less than 80%, 25 ± 1 ℃ of temperature, under the condition of illumination, cultivate, seedling adapts to culture environment after two weeks, removes plastic greenhouse, carries out normal water and fertilizer management (referring to accompanying drawing 6).
Embodiment 2
The present embodiment adopts that embryo callus medium: 2,4-D gets respectively 1.0,1.5mg/L, 2.0,2.5,3.0mg/L; All the other implementation step techniques, with embodiment 1, all can reach the effect of embodiment 1.
Embodiment 3
The present embodiment adopts that embryoid induction medium: 2,4-D gets respectively 0.8,1.0mg/L; All the other implementation step techniques, with embodiment 1, all can reach the effect of embodiment 1.
Embodiment 4
The present embodiment adopts somatic embryo to sprout and seedling medium: GA
3Get respectively 0,0.2,0.8,1.0mg/L, all the other implementation step techniques are with embodiment 1; Perhaps IAA gets respectively 0.1 and 0.25mg/L, and all the other implementation step techniques are with embodiment 1; Perhaps KT gets respectively 0,0.1 and 0.25mg/L, and all the other implementation step techniques, with embodiment 1, all can reach the effect of embodiment 1.
Claims (6)
1. a method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration, is characterized in that, comprises the steps:
1) the taking and sterilizing of explant: get and be in full-bloom stage, length, after the macleaya cordata bud sterilization of 2-12mm, pushes bud aside gently, the stamen of choosing, and the excision filigree, stay flower pesticide as explant, standby;
2) embryo callus induce cultivation: the flower pesticide of step 1) sterilization is inoculated into to BLCG+2,4-D0.5 on～3.0mg/L+6-BA0.5mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000～3000Lx, under the condition of light application time 16～18h/d, carry out inducing of embryo callus, obtain callus after cultivation 30～40d;
3) after callus switching flower pesticide embryoid induction cultivation: by step 2) obtained, form embryoid;
4) somatic embryo is sprouted with seedling and is cultivated: form the whole plant of taking root after the embryoid switching that step 3) is obtained;
5) transplanting of regrowth and cultivation: will take root transplantation of seedlings after hardening in hardening matrix, then carry out normal water and fertilizer management.
2. according to claim 1ly induce macleaya cordata vitro anther monoploid embryo to occur and the method for plant regeneration, it is characterized in that, the process of taking with sterilizing of the described explant of step 1) is specially: get and be in full-bloom stage, length is at the macleaya cordata bud of 2-12mm, with distilled water, clean, clean with aseptic water washing on superclean bench; With 70% alcohol-pickled 10s, then use aseptic water washing 3～4 times after with 0.1% mercuric chloride solution, soaking 8～10min sterilization; Bud is pushed aside gently, the stamen of choosing, the excision filigree, stay flower pesticide as explant, is put in suck dry moisture on the filter paper of aseptic drying, standby.
3. according to claim 1ly induce macleaya cordata vitro anther monoploid embryo to occur and the method for plant regeneration, it is characterized in that, the described flower pesticide embryoid induction of step 3) cultivation process is specially: by step 2) callus that obtains is transferred to MS+2,4-D0.5 on～1mg/L+6-BA0.5mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, carry out the cultivation of inducing of embryoid under the condition of light application time 16-18h/d, form embryoid after 20 days.
4. method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration according to claim 1, is characterized in that, the described somatic embryo of step 4) is sprouted with the seedling cultivation process and is: the embryoid that step 3) is obtained is transferred to MS+GA
3On 0～1mg/L+IAA0～0.25mg/L+KT0～0.25mg/L medium, 25 ± 1 ℃ of temperature, intensity of illumination 1000-3000Lx, cultivate under the condition of light application time 16～18h/d and formed the whole plant of taking root in 10～15 days.
5. method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration according to claim 1, is characterized in that,
The detailed process of step 5) is as follows: the seedling of taking root, be transplanted to peat soil: wood peat soil: in the hardening matrix of coconut palm chaff powder: vermiculite=1:1:1:1, water permeable, and cover seedling with the plastics hut, guarantee that canopy class humidity is not less than 80%, 25 ± 1 ℃ of temperature, intensity of illumination is to cultivate under 3000lx, seedling adapts to culture environment after two weeks, removes plastic greenhouse, carries out normal water and fertilizer management.
6. according to the described method of inducing the generation of macleaya cordata vitro anther monoploid embryo and plant regeneration of claim 1-5 any one, it is characterized in that,
Embryonic callus induction medium: BLCG+2,4-D0.5mg/L+6-BA0.5mg/L;
Flower pesticide embryoid induction medium: MS+2,4-D0.5mg/L+6-BA0.5mg/L;
Somatic embryo is sprouted and seedling medium: MS+GA
30.5mg/L.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105794652A (en) * | 2016-05-23 | 2016-07-27 | 湖南农业大学 | Tissue culture and rapid propagation method of macleaya cordata |
| CN110710452A (en) * | 2019-11-05 | 2020-01-21 | 南京农业大学 | Tissue culture and rapid propagation method of eriobotrya japonica |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1079000A (en) * | 1992-04-01 | 1993-12-01 | 友联坎普公司 | What the taxus somatic embryo took place induces, and produces the alkaloid that contains taxane-ring by it |
| RU2264084C2 (en) * | 2003-12-01 | 2005-11-20 | Федеральное государственное образовательное учреждение высшего профессионального образования Ставропольский государственный аграрный университет | Breeding of plumepoppy cordate (macleaya cordata (willd.) r.br.) (variants) |
-
2013
- 2013-08-15 CN CN201310355266.3A patent/CN103404442B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1079000A (en) * | 1992-04-01 | 1993-12-01 | 友联坎普公司 | What the taxus somatic embryo took place induces, and produces the alkaloid that contains taxane-ring by it |
| RU2264084C2 (en) * | 2003-12-01 | 2005-11-20 | Федеральное государственное образовательное учреждение высшего профессионального образования Ставропольский государственный аграрный университет | Breeding of plumepoppy cordate (macleaya cordata (willd.) r.br.) (variants) |
Non-Patent Citations (2)
| Title |
|---|
| H. KOBLITZ: "Macleaya spp.: Morphogenesis and", 《BIOTECHNOLOGY IN AGRICULTURE AND FORESTRY》, vol. 7, 31 December 1989 (1989-12-31), pages 332 - 348 * |
| H.W.KOHLENBACH: "《Plant Tissue Culture and Its Bio-technological Application》", 31 December 1977, article "Basic Aspects of Differentiation and Plant Regeneration from Cell and Tissue Cultures" * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105794652A (en) * | 2016-05-23 | 2016-07-27 | 湖南农业大学 | Tissue culture and rapid propagation method of macleaya cordata |
| CN105794652B (en) * | 2016-05-23 | 2017-12-01 | 湖南农业大学 | A kind of method of macleaya cordata tissue-culturing quick-propagation |
| CN110710452A (en) * | 2019-11-05 | 2020-01-21 | 南京农业大学 | Tissue culture and rapid propagation method of eriobotrya japonica |
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