CN101869057B - Tissue culture method of floral leaf fatshedera lizei - Google Patents
Tissue culture method of floral leaf fatshedera lizei Download PDFInfo
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- CN101869057B CN101869057B CN2009100498586A CN200910049858A CN101869057B CN 101869057 B CN101869057 B CN 101869057B CN 2009100498586 A CN2009100498586 A CN 2009100498586A CN 200910049858 A CN200910049858 A CN 200910049858A CN 101869057 B CN101869057 B CN 101869057B
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- lizei
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Abstract
The invention relates to a tissue culture method of floral leaf fatshedera lizei, comprising the steps of: obtaining sterile materials, splitting and breeding germ, cultivating adventitious bud strong seedling, cultivating root, hardening seedling, replanting, and the like. Compared with the prior art, the invention greatly improves the breeding speed of floral leaf fatshedera lizei and the uniformity of the seedling, reduces the variation and can realize factory and batch production of grow seedlings.
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 floral leaf fatshedera lizei.
Background technology
Floral leaf fatshedera lizei is an Araliaceae Fatshedrea lizei platymiscium. the evergreenness climbing plant, Gao Keda is more than 1 meter.The nascent stem is herbaceous stem, after lignification gradually.Floral leaf fatshedera lizei is single leaf alternate, and blade is palmate five and splits, and the edge is a white, and petiole is about 8-10cm, and the handle base is the sheath shape and is connected with the stem branch.Adult plant opens light green Xiao Hua in autumn.Floral leaf fatshedera lizei four seasons bule, the extremely strong anti-shady ability of tool suits in the sylvan life mass-planting again.But floral leaf fatshedera lizei becomes different when tissue culture easily, so 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 floral leaf fatshedera lizei of property stability.
The object of the invention can be realized through following technical scheme:
The method for tissue culture of floral leaf fatshedera lizei is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender branch of sprouting spring, remove branches and leaves, 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; 2-4 is visible bud meristematic tissue after week, cultivates 1-2 month again, and little indefinite bud can be grown 3-4cm; Indefinite bud downcut in the proliferated culture medium change bud over to 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 proliferated culture medium of bud, 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 90-95%.
Described axillalry bud inducing culture 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 axillalry bud inducing culture.
The proliferated culture medium of described bud comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/L.
The preferred MS+6-BA1.0mg/L+NAA0.1mg/L of the proliferated culture medium of described bud.
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+NAA0.2mg/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 floral leaf fatshedera lizei and the regularity of seedling through tissue culture technology, has reduced variability, 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 branch of sprouting spring, remove branches and leaves, 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 2 week backs was cultivated 1 month again, and little indefinite bud can be grown 3cm; Indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA0.5mg/L+NAA0.1mg/L over to 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 proliferated culture medium of bud, 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 branch of sprouting spring, remove branches and leaves, 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-BA2.0mg/L+NAA0.2mg/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 3 week backs is cultivated 1 first quarter moon again, and little indefinite bud can be grown 4cm; Indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA1.0mg/L+NAA0.1mg/L over to 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 proliferated culture medium of bud, 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-BA0.5mg/L+NAA0.1mg/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+NAA0.2mg/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 branch of sprouting spring, remove branches and leaves, 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-BA3.0mg/L+NAA0.3mg/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 4 week backs was cultivated 2 months again, and little indefinite bud can be grown 4cm; Indefinite bud downcut in the proliferated culture medium change the bud that comprises MS+6-BA2.0mg/L+NAA0.2mg/L over to 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 proliferated culture medium of bud, 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 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 97%;
(5) refining seedling and transplanting
Culture of rootage 30 days when root system grows to 0.8cm, 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 95%.
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 floral leaf fatshedera lizei is characterized in that, this method may further comprise the steps:
(1) acquisition of sterilizable material
Get the young tender branch of sprouting spring, remove branches and leaves, 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; 2-4 sees the bud meristematic tissue after week, cultivated 1-2 month again, and little indefinite bud is long to 3-4cm; Indefinite bud downcut in the proliferated culture medium change bud over to carry out enrichment culture, 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 proliferated culture medium of bud, 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 90-95%;
Described axillalry bud inducing culture comprises MS+6-BA1.0-3.0mg/L+NAA0.1-0.3mg/L;
The proliferated culture medium of described bud comprises MS+6-BA0.5-2.0mg/L+NAA0.1-0.2mg/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 floral leaf fatshedera lizei according to claim 1 is characterized in that, described axillalry bud inducing culture comprises MS+6-BA2.0mg/L+NAA0.2mg/L.
3. the method for tissue culture of floral leaf fatshedera lizei according to claim 1 is characterized in that, the proliferated culture medium of described bud comprises MS+6-BA1.0mg/L+NAA0.1mg/L.
4. the method for tissue culture of floral leaf fatshedera lizei 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 floral leaf fatshedera lizei according to claim 1 is characterized in that, described root media comprises MS+NAA0.2mg/L.
6. the method for tissue culture of floral leaf fatshedera lizei 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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| CN110537490A (en) * | 2019-09-20 | 2019-12-06 | 上海上房园林植物研究所有限公司 | method for rapidly breeding floral leaf illicium verum discs through tissue culture |
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Non-Patent Citations (3)
| Title |
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| 刘忠荣, 陈屏昭.植物组织培养技术在观赏植物中的应用.《昭通师范高等专科学校学报》.2003,第25卷(第5期),41-43，46. * |
| 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. * |
| 朱锋，等.不同郁闭度香樟林分下熊掌木幼苗生长和生理特性的初步研究.《2008园艺学进展（第八辑）——中国园艺学会第八届青年学术讨论会暨现代园艺论坛论文集》.2008, * |
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