CN101869054B - Tissue culture method for pennisetum setaceum 'Rubrum' - Google Patents
Tissue culture method for pennisetum setaceum 'Rubrum' Download PDFInfo
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- CN101869054B CN101869054B CN2009100498552A CN200910049855A CN101869054B CN 101869054 B CN101869054 B CN 101869054B CN 2009100498552 A CN2009100498552 A CN 2009100498552A CN 200910049855 A CN200910049855 A CN 200910049855A CN 101869054 B CN101869054 B CN 101869054B
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Abstract
The invention relates to a tissue culture method for pennisetum setaceum 'Rubrum'. The method comprises the following steps of: obtaining sterile materials; differentiating and proliferating buds; culturing strong seedlings of adventitious buds; rooting; and hardening off seedlings and transplanting the seedlings and the like. Compared with the prior art, the method greatly improves the reproductive speed of the pennisetum setaceum 'Rubrum' and the trimming of the seedlings, and improves the stability of shape and properties of the pennisetum setaceum 'Rubrum', so that industrial mass production of the seedlings can be realized.
Description
Technical field
The present invention relates to the method for tissue culture of a plant species, especially relate to the method for tissue culture of pennisetum setaceum ' Rubrum '.
Background technology
Pennisetum setaceum ' Rubrum ' is a grass family Pennisetum plant, is distributed in the torrid zone and subtropical zone, is one, the biennial herb plant, plant height 80cm, and the leaf consor is in base portion, and blade is flat, and it is linear to be purple, and it is coniform that inflorescence is.Pennisetum setaceum ' Rubrum ' is the external famous grass of viewing and admiring; Can be used for afforestation or bank; The colored fringe of pennisetum setaceum ' Rubrum ' leaf is purple, and the garden cultivation is grand and graceful, can also spend the border, view and admire and use in the careless garden belonging to a category; Have good annidation and ornamental value, be good presbyopic glasses with ground by material.But as the new varieties of external introduction, pennisetum setaceum ' Rubrum ' is introduced a fine variety negligible amounts, and the seedling supply receives certain restriction.
Summary of the invention
The object of the invention is exactly for the defective that overcomes above-mentioned prior art existence a kind of regularity that improves reproduction speed and seedling to be provided, and improves the method for tissue culture of the pennisetum setaceum ' Rubrum ' of property stability.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of pennisetum setaceum ' Rubrum ' is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, with behind the running water flushing 1-3h on superclean bench; Utilize the alcohol immersion 10-50s of mass concentration successively for 70-75%; Volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, uses aseptic water washing 4-6 time again, utilize aseptic filter paper to blot surperficial moisture after; Be cut into the sections of the long band axillalry bud of 0.5-2cm again, sections is inoculated on the axillalry bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the axillalry bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 3-5 is visible bud meristematic tissue after week, cultivates 1-2 month again, and little indefinite bud can be grown 3-4cm; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium, the callus of indefinite bud base portion is more, but does not influence propagation; Indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has the 2-3 strain to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base and grows, and indefinite bud extends rapidly, can grow 3-4cm after 15-30 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3-4cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 5-15 days, can grow to 4-6cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
Culture of rootage 15-30 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 2-4 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 20-40 days; Can transplant outdoorly, give rich water quality management, final transplanting survival rate is 90-100%.
Described axillalry bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L.
The preferred MS+6-BA5.0mg/L+NAA0.5mg/L of described axillalry bud inducing culture.
Described adventitious bud proliferation medium comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L.
The preferred MS+6-BA2.0mg/L+NAA0.2mg/L of described adventitious bud proliferation medium.
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA0.5mg/L+NAA0.1mg/L of described strong seedling culture base.
Described root media comprises MS+NAA0.1-0.3mg/L.
The preferred MS+NAA0.1mg/L of described root media.
Described medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx.
Compared with prior art, the present invention has greatly improved the reproduction speed of pennisetum setaceum ' Rubrum ' and the regularity of seedling, and has improved the stability of its proterties through tissue culture technology, can realize the batch production production in enormous quantities of growing seedlings.
Embodiment
Below in conjunction with specific embodiment the present invention is elaborated.
Embodiment 1
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, with behind the running water flushing 1h on superclean bench; Utilizing mass concentration successively is 70% alcohol immersion 10s; Volumetric concentration is that 0.5 ‰ mercury soaks 10min, uses aseptic water washing again 4 times, utilize aseptic filter paper to blot the moisture on surface after; Be cut into the sections of the long band axillalry bud of 0.5cm again, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA1.0mg/L+NAA0.1mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 1 week, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 3 week backs was cultivated 1 month again, and little indefinite bud can be grown 3cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA1.0mg/L+NAA0.1mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation; Indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has 2 strains to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state; After the bud of will growing thickly is divided into Xiao Cong; Go on the strong seedling culture base that comprises MS+6-BA0.5mg/L+NAA0.1mg/L and grow, indefinite bud extends rapidly, can grow 3cm after 15 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3cm, root induction in the root media of going into to comprise MS+NAA0.1mg/L of transferring, the seedling base section dissolves the root original hase of white after 5 days, can grow to 4cm after 25 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90%;
(5) refining seedling and transplanting
Culture of rootage 15 days when root system grows to 0.5cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 2 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 20 days; Can transplant outdoorly, give rich water quality management, final transplanting survival rate is 90%.
The medium of above-mentioned various situation also comprises sucrose 20g/L, agar powder 4g/L, pH=5.5,24 ℃ of cultivation temperature, illumination 1500lx.
Embodiment 2
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, with behind the running water flushing 2h on superclean bench; Utilizing mass concentration successively is 75% alcohol immersion 30s; Volumetric concentration is that 1 ‰ mercury soaks 15min, uses aseptic water washing again 5 times, utilize aseptic filter paper to blot the moisture on surface after; Be cut into the sections of the long band axillalry bud of 1cm again, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 2 weeks, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 4 week backs was cultivated 1 month again, and little indefinite bud can be grown 4cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA2.0mg/L+NAA0.2mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation; Indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has 3 strains to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state; After the bud of will growing thickly is divided into Xiao Cong; Go on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L and grow, indefinite bud extends rapidly, can grow 4cm after 20 days;
(4) culture of rootage
Get the indefinite bud plantlet of 4cm, root induction in the root media of going into to comprise MS+NAA0.3mg/L of transferring, the seedling base section dissolves the root original hase of white after 10 days, can grow to 6cm after 30 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 100%;
(5) refining seedling and transplanting
Culture of rootage 20 days when root system grows to 1cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 3 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 30 days; Can transplant outdoorly, give rich water quality management, final transplanting survival rate is 100%.
The medium of above-mentioned various situation also comprises sucrose 30g/L, agar powder 6g/L, pH=5.8,25 ℃ of cultivation temperature, illumination 2000lx.
Embodiment 3
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, with behind the running water flushing 3h on superclean bench; Utilizing mass concentration successively is 75% alcohol immersion 50s; Volumetric concentration is that 2 ‰ mercury soaks 30min, uses aseptic water washing again 6 times, utilize aseptic filter paper to blot the moisture on surface after; Be cut into the sections of the long band axillalry bud of 2cm again, sections is inoculated on the axillalry bud inducing culture that comprises MS+6-BA5.0mg/L+NAA0.5mg/L;
(2) differentiation of bud and propagation
Sections was inoculated on the axillalry bud inducing culture after 3 weeks, and the axillalry bud position begins to expand, and green projection occurs; The visible bud meristematic tissue in 5 week backs was cultivated 2 months again, and little indefinite bud can be grown 4cm; Indefinite bud downcut to change in the adventitious bud proliferation medium that comprises MS+6-BA3.0mg/L+NAA0.3mg/L carry out enrichment culture, the callus of indefinite bud base portion is more, but does not influence propagation; Indefinite bud growth rapidly and do not have bad phenomenon such as vitrifying, it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium; Every clump has 3 strains to extend in the bud of growing thickly that induces; Remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base that comprises MS+6-BA2.0mg/L+NAA0.2mg/L and grows; Indefinite bud extends rapidly, can grow 3.5cm after 30 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3.5cm, root induction in the root media of going into to comprise MS+NAA0.3mg/L of transferring, the seedling base section dissolves the root original hase of white after 15 days, can grow to 5cm after 40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 91%;
(5) refining seedling and transplanting
Culture of rootage 30 days when root system grows to 0.7cm, is selected the aseptic seedling of well developed root system, robust growth; Refine seedling 4 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 40 days; Can transplant outdoorly, give rich water quality management, final transplanting survival rate is 94%.
The medium of above-mentioned various situation also comprises sucrose 40g/L, agar powder 8g/L, pH=6.0,26 ℃ of cultivation temperature, illumination 2500lx.
Claims (6)
1. the method for tissue culture of pennisetum setaceum ' Rubrum ' is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender bud of sprouting spring, remove the blade of coated outside after, with behind the running water flushing 1-3h on superclean bench; Utilize the alcohol immersion 10-50s of mass concentration successively for 70-75%; Volumetric concentration is that the mercury of 0.5-2 ‰ soaks 10-30min, uses aseptic water washing 4-6 time again, utilize aseptic filter paper to blot surperficial moisture after; Be cut into the sections of the long band axillalry bud of 0.5-2cm again, sections is inoculated on the axillalry bud inducing culture;
(2) differentiation of bud and propagation
Sections is inoculated in that 1-3 is after week on the axillalry bud inducing culture, and the axillalry bud position begins to expand, and green projection occurs; 3-5 sees the bud meristematic tissue after week, cultivated 1-2 month again, and little indefinite bud is long to 3-4cm; Indefinite bud downcut to change over to carry out enrichment culture in the adventitious bud proliferation medium, the callus of indefinite bud base portion is more, but does not influence propagation; The indefinite bud growth is rapid and do not have the vitrifying bad phenomenon, and it is good in medium, to grow;
(3) indefinite bud strong seedling culture
On the adventitious bud proliferation medium, every clump has the 2-3 strain to extend in the bud of growing thickly that induces, and remaining is in the dwarfing state, after the bud of will growing thickly is divided into Xiao Cong, goes on the strong seedling culture base and grows, and indefinite bud extends rapidly, long to 3-4cm after 15-30 days;
(4) culture of rootage
Get the indefinite bud plantlet of 3-4cm, root induction in the root media is gone in switching, and the seedling base section dissolves the root original hase of white after 5-15 days, grows to 4-6cm after 25-40 days, and root system is sturdy, and fibrous root is numerous, and rooting rate is 90-100%;
(5) refining seedling and transplanting
Culture of rootage 15-30 days, when root system grows to 0.5-1cm, select the aseptic seedling of well developed root system, robust growth; Refine seedling 2-4 days in indoor uncork, then seedling is taken out, clean the agar of root; Plant in the seedbed in the greenhouse and tamed 20-40 days; Promptly transplant outdoorly, give rich water quality management, final transplanting survival rate is 90-100%;
Described axillalry bud inducing culture comprises MS+6-BA1.0-5.0mg/L+NAA0.1-0.5mg/L;
Described adventitious bud proliferation medium comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L;
Described strong seedling culture base comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L;
Described root media comprises MS+NAA0.1-0.3mg/L.
2. the method for tissue culture of pennisetum setaceum ' Rubrum ' according to claim 1 is characterized in that, described axillalry bud inducing culture comprises MS+6-BA5.0mg/L+NAA0.5mg/L.
3. the method for tissue culture of pennisetum setaceum ' Rubrum ' according to claim 1 is characterized in that, described adventitious bud proliferation medium comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
4. the method for tissue culture of pennisetum setaceum ' Rubrum ' according to claim 1 is characterized in that, described strong seedling culture base comprises MS+6-BA0.5mg/L+NAA0.1mg/L.
5. the method for tissue culture of pennisetum setaceum ' Rubrum ' according to claim 1 is characterized in that, described root media comprises MS+NAA0.1mg/L.
6. the method for tissue culture of pennisetum setaceum ' Rubrum ' according to claim 1 is characterized in that, each medium also comprises sucrose 20-40g/L, agar powder 4-8g/L, medium pH 5.5-6.0, cultivation temperature 24-26 ℃, illumination 1500-2500lx respectively.
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| CN104719158A (en) * | 2015-03-09 | 2015-06-24 | 中国农业科学院兰州畜牧与兽药研究所 | Method for rapidly establishing medium-sized Chinese pennisetum herb tissue culture regeneration system by taking seeds as explants |
| CN106148400A (en) * | 2016-09-09 | 2016-11-23 | 福建农林大学 | A kind of Agrobacterium tumefaciens mediated tomato conversion method |
| CN106332781B (en) * | 2016-09-29 | 2018-07-13 | 遵义市龙驰生物科技有限公司 | A kind of efficient expanding propagation method of hybrid Chinese pennisetum |
| CN110692518A (en) * | 2019-10-11 | 2020-01-17 | 高州市石生源生物科技发展有限公司 | Tissue culture rapid propagation breeding method for Guangdong grass |
| CN112655556A (en) * | 2020-12-22 | 2021-04-16 | 中国热带农业科学院热带作物品种资源研究所 | Tissue culture detoxification and rapid propagation method of wangcao |
| CN115362939B (en) * | 2022-10-27 | 2023-01-24 | 西南林业大学 | Tissue culture method and application of pennisetum setosum |
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Non-Patent Citations (2)
| Title |
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| 唐道城,梁顺祥.观赏植物组织培养研究进展.《青海大学学报( 自然科学版)》.2006,第24卷(第4期),5-9. * |
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Owner name: SHANGHAI SHANGFANG LANDSCAPE PLANT RESEARCH INSTIT Free format text: FORMER OWNER: SHANGHAI SHANGFANG LANDSCAPE PLANT INSTITUTE Effective date: 20140102 Owner name: STATE GRID SHANGHAI ELECTRIC POWER COMPANY POWER S Effective date: 20140102 |
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