CN103931497A - Method for improving seedling rate of tissue culture seedlings of hylocereus undulatus britt - Google Patents

Method for improving seedling rate of tissue culture seedlings of hylocereus undulatus britt Download PDF

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CN103931497A
CN103931497A CN201410135613.6A CN201410135613A CN103931497A CN 103931497 A CN103931497 A CN 103931497A CN 201410135613 A CN201410135613 A CN 201410135613A CN 103931497 A CN103931497 A CN 103931497A
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seedling
adventitious bud
root
roots
stem
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CN103931497B (en
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彭绿春
瞿素萍
张艺萍
苏艳
王丽花
杨秀梅
张丽芳
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Flower Research Institute of YAAS
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Abstract

The invention provides a method for improving the seedling rate of tissue culture seedlings of hylocereus undulatus britt. The method comprises the following steps: carrying out adventitious bud induction, synchronous multiplication and rooting seedling culture, wherein hylocereus undulatus britt barbed seats and lignification shoots with small barbs and fluff on the barbed seats are used as explants; when lateral branches of adventitious buds grow, base roots of adventitious bud base parts grow and aerial roots of the lateral branches grow, carrying out hardening seedling; after the hardening seedling, taking out seedlings from a bottle; cutting off the lateral branches with the aerial roots and main stems with the base roots and planting in a culture medium; after the lateral branches and the main stems are survived, growing new roots; and putting the lateral branches and the main stems into pots. According to the method, the production period of the tissue culture seedlings of the hylocereus undulatus britt is short, the adventitious bud induction rate is high, the multiplication and the rooting are synchronous, the multiplication coefficient is more than 5.45 and the rooting rate is 100%; the growth vigor of multiplied seedlings is strong and the transplanting survival rate is more than 95%; one inoculated adventitious bud can grow 3-4 lateral branches with the aerial roots, namely one inoculated adventitious bud can grow 4-5 grown-up seedlings with the roots; the seedling rate of the tissue culture seedlings of the hylocereus undulatus britt is increased by 3-4 times.

Description

A kind of method that improves dragon fruit group training seedling planting percent
Technical field
The invention belongs to plant propagation technical field, be specifically related to a kind of method of the planting percent that improves dragon fruit group training seedling.
Background technology
Dragon fruit formal name used at school hylocereus ( hylocereus undatus(Haw.) Britt. & Rose.), have another name called red dragon fruit, Green Dragon fruit, celestial mamoncillo etc., be that Cactaceae (Cactaceae) hylocereus belongs to (Hylocereus) plant.Not only good looking appearance of the fruit of dragon fruit, and with health role, there is purifying the blood, fall effect of fiery norcholesterol, normal food can promote human body metabolism, improves immunologic function, and its cultivation is comparatively easy in addition, damage by disease and insect is few, early fruiting and high yield, can adapt to arid climate, is kind of a novel fruits that has market development potential.In recent years, the industries such as the plantation of dragon fruit, processing, tourist sight are all being greatly developed in the province such as Yunnan, Guizhou, Guangxi, Guangdong.Under this background, dragon fruit seedling market is fiery, and supply falls short of demand for seedling.
At present, the mode that dragon fruit seedling is produced has seminal propagation, cottage propagation and tissue culture propagation, and wherein tissue cultivation is the effective way that Fast-propagation is produced dragon fruit seedling.Existing dragon fruit group culturation rapid propagating technology is generally taking stems of dragon fruits section as explant, through adventitious bud inducing, breed and three steps of taking root complete plant regeneration.Exist that the basic element of cell division working concentration of propagation in incubation is too high and the whole production cycle is long (as conventional three-step approach regeneration plant from explant bottle inlet to bottle transplantation of seedlings the needs time of at least 110 days) two large shortcomings; the problem such as the seedling aberration rate of the production causing is thus high, quality is unstable, production cost is high; seriously reduce dragon fruit group and cultivated seedling industrialized production efficiency, greatly limited the scale of its production.Also have propagation and take root and synchronously carry out, but the group that these tissue culture methods are produced training seedling only has base root, there is no aerial root (aerial root refers to the root growing on the newly-increased side shoot of group training seedling), a strain group training seedling only has base root, the finished product seedling of group training seedling and formation is man-to-man, and a strain group training seedling can only form at most a strain finished product seedling, therefore, planting percent is low, and production cost is higher.
Summary of the invention
The technical problem to be solved in the present invention is to overcome lower defect and the deficiency of existing dragon fruit group training seedling planting percent, to improve the planting percent of dragon fruit group training seedling, more effectively reduce the production cost of seedling, a kind of method that can improve dragon fruit group training seedling planting percent is provided.The method comprises the following steps:
(1) adventitious bud inducing
1. the selection of explant and processing: select semi-lignified, without the stems of dragon fruits section of damage by disease and insect, first clean with washing powder water, and after rinsing well with running water, this stem section is longitudinally cut, remove marrow, being cut into wide is 2cm~3cm, the long stem sheet for 3cm~4cm again, and the crust of every stem tuber sheet all stings seat with one, and retains fine hair and spinule on thorn seat;
2. adventitious bud inducing: the stem sheet of handling well is inoculated on adventitious bud induction culture base after sterilizing, cutting upper and lower two end surfaces, described adventitious bud induction culture base is MS+TDZ0.4~0.6mg/L+NAA0.1 mg/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8;
(2) seedling is cultivated
When indefinite bud grows to 3~5cm length, cut indefinite bud, indefinite bud is accessed in seedling medium, or indefinite bud is divided in two sections of access seedling medium, described seedling medium is MS+6-BA 2~4 mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8;
Step (1) 2. and the condition of culture of step (2) be: temperature is 25 DEG C~28 DEG C, and intensity of illumination is 3000lx~5000lx, and light application time is 14 h/d, and in each medium, the percentage of sucrose and agar content is mass percent;
(3) acclimatization and transplants
Seedling is cultured to and on indefinite bud, grows side shoot, and when indefinite bud base portion grows Ji Gen and side shoot and grows aerial root, bottle seedling is moved in the booth of outdoor ventilation, hardening 6~7 days, two days cover the shading screen of 50% shading rate, hardening finishes, seedling is taken out in bottle, clean medium, with sharp and through the pocket knife of alcohol disinfecting, cut with the side shoot of aerial root and with the stem of base root, be put in 1000 times of liquid of 75% tpn wetting powder and soak 10 minutes, taking-up is placed on shady and cool place and dries in the air 1~2 hour, plant afterwards in filling the square position of matrix, seeding row spacing is 2cm × 4cm, matrix in square position is perlite, in advance perlite is watered permeable, after plantation, square position is positioned in booth and is managed, within 3~5 days, water water one time, keep 22 DEG C~25 DEG C of canopy temperatures, relative air humidity 50~60%, surviving and growing new root with the side shoot of aerial root and with the stem of base root in square position to be planted in, move on to maintenance in pot for growing seedlings.
Step (1) 2. preferred adventitious bud induction culture base is: MS+TDZ 0.4mg/L+NAA0.1 mg/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8; The preferred seedling medium of step (2) is: MS+6-BA 4mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, pH value is 5.8.
compared with prior art, the invention has the beneficial effects as follows:
The inventive method adopts special explant processing, the hormone of low concentration and the coordinated of each step, produce the effect of Synergistic, not only make dragon fruit group cultivate the with short production cycle of seedling, adventitious bud induction frequency high (more than 80%), propagation is synchronizeed with taking root, growth coefficient reaches more than 5.45, propagation seedling growing way is vigorous, rooting rate can reach 100%, group training seedling becomes the survival rate of transplantation of seedlings to reach more than 95%, and an indefinite bud of inoculation can grow 3~4 side shoots with aerial root, side shoot is sturdy, can make an indefinite bud of inoculation breed the seedling (the stem section of 3~4 side shoots with aerial root and a strain tape base root is the seedling of 4~5 strain band roots and organizes training seedling seedling) of 4~5 strain band roots, make group training seedling planting percent improve 300~400%, the productivity ratio that is dragon fruit group seedling seedling has improved 3~4 times, production efficiency is greatly improved, reduce significantly production cost, compared with prior art produce unforeseeable technique effect.
Embodiment
A kind of method that improves dragon fruit group training seedling planting percent of embodiment 1 the present invention, its step is as follows:
(1) adventitious bud inducing
1. the selection of explant and processing: select semi-lignified, without the stems of dragon fruits section of damage by disease and insect, first clean with washing powder water, and after rinsing well with running water, this stem section is longitudinally cut, remove marrow, be cut into widely for 2cm~3 cm, length are the stem sheet of 3 cm~4cm, the crust of every stem tuber sheet is all with a complete thorn seat again, and retains fine hair and spinule in thorn.
2. adventitious bud inducing: 30s sterilizes in the alcohol that is first 75% at mass fraction by the stem sheet of handling well, in sterile water, clean after 1 time, proceed to mass fraction and be the 10min that sterilizes in 2% liquor natrii hypochloritis, sterile water wash 3~5 times afterwards, after having sterilized, be inoculated on adventitious bud induction culture base after cutting upper and lower two end surfaces, described adventitious bud induction culture base is: MS+TDZ 0.6mg/L+NAA 0.1 mg/L+ sucrose 3%+ agar 0.6%, pH value is 5.8, and adventitious bud induction frequency is 80%;
(2) seedling is cultivated (synchronously breed and take root)
After 35 days~40 days, indefinite bud goes out from the thorn seat director of stem sheet, when indefinite bud grows to 3~5cm length, cut indefinite bud, shorter indefinite bud directly accesses in seedling medium, long is divided into indefinite bud in two sections of access seedling medium (increasing indefinite bud quantity), and described seedling medium is MS+6-BA 2mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8;
Step (1) 2. and the condition of culture of step (2) be: temperature is 25 DEG C~28 DEG C, and intensity of illumination is 3000lx~5000lx, and light application time is 14 h/d, and in each medium, the percentage of sucrose and agar content is mass percent;
(3) acclimatization and transplants
Seedling was cultivated after 30 days, and indefinite bud grows the overgrowing branch that length is 3~4cm, and indefinite bud base portion grows Ji Gen and side shoot grows aerial root, now, bottle seedling is moved in the booth of outdoor ventilation to hardening 6~7 days, two days cover the shading screen of 50% shading rate, throw off afterwards described shading screen.Hardening finishes, seedling is taken out in bottle, clean medium, with sharp and through the pocket knife of alcohol disinfecting, cut with the side shoot of aerial root and with the stem of base root, be put in 1000 times of liquid of 75% tpn wetting powder and soak 10 minutes, taking-up is placed on shady and cool place and dries in the air 1~2 hour, plant afterwards in filling the square position of matrix, seeding row spacing is 2cm × 4cm, again square position is positioned in booth and is managed, within 3~5 days, water water one time, keep 22 DEG C~25 DEG C of canopy temperatures, relative air humidity 50~60%, surviving and growing new root with the side shoot of aerial root and with the stem of base root in square position to be planted in, move on to maintenance in pot for growing seedlings, matrix in square position is perlite, before plantation, waters in advance permeable to perlite.After approximately 30 days, seedling survives and grows new root.
embodiment 2
Embodiment 2 is except following measures difference, and all the other measures are identical with embodiment 1, repeat no more.
Step (1) adventitious bud induction culture base is 2.: MS+TDZ 0.4mg/L+NAA0.1 mg/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8; The seedling medium of step (2) is: MS+6-BA 4mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, pH value is 5.8.
the test effect of each embodiment above:
Dragon fruit group training seedling seedling: be refer to an access adventitious bud proliferation (growing) in above-mentioned seedling medium with the stem of base root and/or with the side shoot of aerial root.
These have root, stem, leaf (spinule of dragon fruit is its leaf, namely abnormal leaf) with the stem of base root and with the side shoot of aerial root, and after cutting respectively, becoming complete regenerated plant is seedling.
Dragon fruit group training seedling planting percent: be the ratio that refers to an access the number of branch with aerial root of an adventitious bud proliferation in seedling medium and the stem with base root of this indefinite bud and count sum and this indefinite bud number, its computing formula is:
Each embodiment accesses the indefinite bud in seedling medium above, its base portion grows a large amount of base roots, an indefinite bud can grow 3~4 side shoots with aerial root, every side shoot grows 2~4 aerial roots, therefore, an indefinite bud can cut 3~4 side shoots with aerial root and a stem with base root, therefore an indefinite bud can breed 4~5 strains of group training seedling seedling, group training planting percent is 400~500%, and in prior art, inoculate an indefinite bud and can not produce the side shoot with aerial root, indefinite bud can only breed the group training seedling seedling of a strain tape base root, therefore, the inventive method can make dragon fruit group training seedling planting percent improve 300~400%, production efficiency is greatly improved, reduce significantly production cost, compared with prior art produce unforeseeable technique effect.
Each embodiment is because the group training seedling stem base portion of breeding grows also more (every side shoot grows 2~4 aerial roots) of aerial root on a large amount of base roots, side shoot above, its propagation seedling growing way is vigorous, side shoot is sturdy, therefore, cut with the side shoot of aerial root and to be transplanted to the survival rate of matrix with the group training seedling seedling of the stem of base root also high, transplanting survival rate is more than 95%, survive soon, within 30~35 days, can survive and grow new root.
Each embodiment dragon fruit group is cultivated the with short production cycle of seedling above: cultivate (synchronously breed and take root) (30 days~35 days) from adventitious bud inducing (35 days~40 days) to seedling and only need 65~75 days, form complete regenerated plant and only need 65~75 days, survive and need 30 days~35 days through acclimatization and transplants again, being induced to acclimatization and transplants from indefinite bud only survives and to need 95 days~110 days, therefore, dragon fruit group is cultivated the with short production cycle of seedling.
Each embodiment adopts special explant processing above, the hormone of low concentration and the coordinated of each step, produce the effect of Synergistic, not only make dragon fruit group cultivate the with short production cycle of seedling, adventitious bud induction frequency high (more than 80%), propagation is synchronizeed with taking root, growth coefficient reaches more than 5.45, rooting rate can reach 100%(in table 1), propagation seedling growing way is vigorous, group training seedling seedling transplanting survival rate reaches more than 95%, and an indefinite bud of inoculation can grow 3~4 more side shoots with aerial root, side shoot is sturdy, can make an indefinite bud of inoculation breed the seedling (i.e. group training seedling seedling) of 4~5 strain band roots, make group training seedling planting percent improve 300~400%, the productivity ratio of dragon fruit group training seedling seedling has improved 3~4 times, production efficiency is greatly improved, low significantly production cost, compared with prior art produce unforeseeable technique effect.

Claims (2)

1. a method that improves dragon fruit group training seedling planting percent, comprises the following steps:
(1) adventitious bud inducing
1. the selection of explant and processing: select semi-lignified, without the stems of dragon fruits section of damage by disease and insect, first clean with washing powder water, and after rinsing well with running water, this stem section is longitudinally cut, remove marrow, be cut into widely for 2cm~3 cm, length are the stem sheet of 3 cm~4cm, the crust of every stem tuber sheet is all with a thorn seat again, and retains fine hair and spinule in thorn;
2. adventitious bud inducing: the stem sheet of handling well is inoculated on adventitious bud induction culture base after sterilizing, cutting upper and lower two end surfaces, described adventitious bud induction culture base is MS+TDZ 0.4~0.6mg/L+NAA 0.1 mg/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8;
(2) seedling is cultivated
When indefinite bud grows to 3~5cm length, cut indefinite bud, indefinite bud is accessed in seedling medium, or indefinite bud is divided in two sections of access seedling medium, described seedling medium is MS+6-BA 2~4 mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, and pH value is 5.8;
Step (1) 2. and the condition of culture of step (2) be: temperature is 25 DEG C~28 DEG C, and intensity of illumination is 3000~5000lx, and light application time is 14 h/d, and in each medium, the percentage of sucrose and agar content is mass percent;
(3) acclimatization and transplants
Seedling is cultured to and on indefinite bud, grows side shoot, and when indefinite bud base portion grows Ji Gen and side shoot and grows aerial root, bottle seedling is moved in the booth of outdoor ventilation, hardening 6~7 days, two days cover the shading screen of 50% shading rate, hardening finishes, seedling is taken out in bottle, clean medium, with cutting through the pocket knife of alcohol disinfecting with the side shoot of aerial root and with the stem of base root, be put in 1000 times of liquid of 75% tpn wetting powder and soak 10 minutes, taking-up is placed on shady and cool place and dries in the air 1~2 hour, plant afterwards in filling the square position of matrix, seeding row spacing is 2cm × 4cm, matrix in square position is perlite, before plantation, water in advance permeable to perlite, after plantation, square position is positioned in booth and is managed, within 3~5 days, water water one time, keep 22 DEG C~25 DEG C of canopy temperatures, relative air humidity 50~60%, surviving and growing new root with the side shoot of aerial root and with the stem of base root in square position to be planted in, move on to maintenance in pot for growing seedlings.
2. a kind of method that improves dragon fruit group training seedling planting percent according to claim 1, step (1) adventitious bud induction culture base is 2.: MS+TDZ 0.4mg/L+NAA0.1 mg/L+ sucrose 3%+ agar 0.6%, pH value is 5.8; The seedling medium of step (2) is: MS+6-BA 4mg/L+NAA 0.1 mg/L+ active carbon 1g/L+ sucrose 3%+ agar 0.6%, pH value is 5.8.
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CN107347494A (en) * 2017-08-08 2017-11-17 南农新农村发展研究院句容有限公司 A kind of method of dragon fruit nursery Rapid Rooting
CN108040881A (en) * 2017-12-28 2018-05-18 广西南宁桂尔创环保科技有限公司 A kind of dragon fruit synchronization tissue culture culture medium and the cultural method of plant
CN108770695A (en) * 2018-09-10 2018-11-09 西南林业大学 A kind of method of the in vitro tissue culture sprouting and rooting of red heart dragon fruit
CN108812318A (en) * 2018-06-29 2018-11-16 钦州市农业科学研究所 Hylocereus undatus tissue-culturing rapid propagation culture medium
CN108849506A (en) * 2018-06-29 2018-11-23 广西驰胜农业科技有限公司 A kind of method of dragon fruit tissue cultivating and seedling
CN109042332A (en) * 2018-09-13 2018-12-21 四川七彩林业开发有限公司 A kind of tissue culture and rapid propagation method of tripterygium wilfordii
CN113229146A (en) * 2021-06-02 2021-08-10 中国热带农业科学院橡胶研究所 Tissue culture, transplantation and domestication method for red-meat pitaya
CN114128503A (en) * 2022-01-17 2022-03-04 山西农业大学 Soilless rooting cutting propagation method for sedum
CN114279780A (en) * 2021-12-07 2022-04-05 海南大学 Chromosome and karyotype analysis method using pitaya stem tips as materials

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Cited By (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN107347494A (en) * 2017-08-08 2017-11-17 南农新农村发展研究院句容有限公司 A kind of method of dragon fruit nursery Rapid Rooting
CN108040881A (en) * 2017-12-28 2018-05-18 广西南宁桂尔创环保科技有限公司 A kind of dragon fruit synchronization tissue culture culture medium and the cultural method of plant
CN108812318A (en) * 2018-06-29 2018-11-16 钦州市农业科学研究所 Hylocereus undatus tissue-culturing rapid propagation culture medium
CN108849506A (en) * 2018-06-29 2018-11-23 广西驰胜农业科技有限公司 A kind of method of dragon fruit tissue cultivating and seedling
CN108770695A (en) * 2018-09-10 2018-11-09 西南林业大学 A kind of method of the in vitro tissue culture sprouting and rooting of red heart dragon fruit
CN108770695B (en) * 2018-09-10 2022-01-21 西南林业大学 Method for in-vitro tissue culture and rapid seedling establishment of red-heart pitaya
CN109042332A (en) * 2018-09-13 2018-12-21 四川七彩林业开发有限公司 A kind of tissue culture and rapid propagation method of tripterygium wilfordii
CN113229146A (en) * 2021-06-02 2021-08-10 中国热带农业科学院橡胶研究所 Tissue culture, transplantation and domestication method for red-meat pitaya
CN114279780A (en) * 2021-12-07 2022-04-05 海南大学 Chromosome and karyotype analysis method using pitaya stem tips as materials
CN114279780B (en) * 2021-12-07 2023-12-12 海南大学 Chromosome and karyotype analysis method using dragon fruit stem tip as material
CN114128503A (en) * 2022-01-17 2022-03-04 山西农业大学 Soilless rooting cutting propagation method for sedum

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