CN108419675B - Tissue culture seedling raising method for passion fruit top shoots - Google Patents
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Abstract
A tissue culture seedling method of passion fruit top shoot comprises the technical links of explant browning resistance, disinfection treatment, axillary bud initial induction culture, callus induction culture, subculture proliferation culture, rooting induction culture and bottle seedling transplantation; the young tender tips are taken as explants, are soaked in a mixed solution of an antioxidant Vc and citric acid, and are soaked in a potassium permanganate solution for sterilization, so that the oxidation resistance and browning resistance and activity of the young explants are improved; adding active carbon and polyvinylpyrrolidone into culture medium of 3 stages of axillary bud primary induction, callus induction and subculture proliferation for adsorbing secreted harmful substances and inhibiting browning substance generation. By adopting the method, the initial induced pollution rate and browning rate of the passion flower tips can be controlled to be below 10 percent; the initial induction survival rate of the top shoot reaches more than 90 percent, the subculture multiplication coefficient of the cluster buds is more than 5.0, and the rooting rate is more than 90 percent; the survival rate of bottle seedling transplantation reaches more than 95.0 percent. The invention can meet the requirement of industrial seedling culture.
Description
Technical Field
The invention relates to a crop cultivation technology, in particular to a tissue culture seedling raising method for passion fruit top shoots.
Background
Passiflora caerulea L, a perennial evergreen climbing woody vine, is an aromatic and delicious fruit with the reputation of "the king of fruit juice". The fruit contains rich trace elements such as calcium, iron, phosphorus and the like and 17 amino acids and other substances, and has good health-care and medicinal effects besides eating. It has been determined that passion fruit contains over 135 aromatic substances and is most suitable for processing into fruit juice. However, the diseases of the planted passion fruit are serious, mainly include root rot, phytophthora root rot, anthracnose, mosaic disease and the like, and particularly in perennial orchards, the diseases are more serious, so that the number of dropped fruits and defective fruits is increased, and the yield and the quality of the orchards are seriously influenced. Researches show that passion flower diseases are mainly related to passion flower mosaic virus CMV and lignification virus PWV carried by seedlings, and the best solution is to culture virus-free seedlings by adopting a tissue culture mode.
Tissue culture seedling raising of passion flower has been reported more. The tissue culture of the species of passion flower, common cork passion flower, stinky passion flower, passion fruit flower and the like is mainly carried out by taking leaves as explants abroad, and most of the tissue culture takes MS as a basic culture medium. The research shows that: the induced differentiation of different species varies under the same hormone formulation, e.g. 1mg.L-1Cytokinin 6-BA and 1mg.L-1After the combination of the auxin NAA, the passion flower and the wood bolt passion flower can form buds and tips in the test tube, but the stink passion flower only grows roots, and the yellow fruit passion flower only generates callus. Chengzhiying et al (1986) also used MS culture medium to perform tissue culture on the passion fruit leaves and stems, and the research shows that: the cluster bud differentiation is only 2-4 mg.L-16-BA and 0.4mg.L-1The IAA combination shows that the differentiation rate is only 5-20%; the concentration of 6-BA is reduced to 0.5mg.L-1After the method is adopted, the bud clumps grow high to form rootless seedlings; the optimal root induction culture medium is 1/8MS, and the addition concentration is 0.2mg.L-1IAA and NAA, the rooting rate is 67.0%. Wegener et al (CN 1O579464 OA) disclose a tissue culture seedling method of stem segments of gold passion fruits, which considers that 0.05-0.35 mg.L of gold passion fruits are added into an MS culture medium-1IAA、12～22 mg.L-1Leucine and 1.5-3.5 g.L-1After glycerol is added, the induction culture of the cluster buds is facilitated; adding 0.2-0.55 mg.L into MS culture medium-1IAA、0.5～3.0 mg.L-1NAA、15～50 mg.L-1Leucine and 2.5-5.0 g.L-1After glycerol, the subculture multiplication culture of the cluster buds is facilitated; adding 1.2-4.5 mg.L into MS culture medium-1IAA、1.5～4.0 mg.L-1NAA、35～80 mg.L-1Leucine and 8-12 g.L-1And the glycerol is beneficial to hardening and culturing the rootless seedlings. Mushroom fungus bags, organic fertilizer and air-dried pond sludge in a weight ratio of 2: 1.5: 3 are used as a matrix, and the non-rooted seedlings after seedling strengthening are taken out of the bottle to root. Robles (1978) performed test tube culture of axillary buds of Passiflora incarnata and Passiflora incarnata, and the results showed that Nitsch, Blaydes and MS culture media all induced axillary bud differentiation of callus, and 2mg.L of the culture media was added-1The Kt effect is the best. Zhang Yu Wei (CN 106106175A)
It is considered that 0.25 to 0.3mg.L of the culture medium for MS is added-1NAA and 0.35-0.5 mg.L-16-BA, which is favorable for axillary bud differentiation into adventitious bud, 0.25-0.3 mg.L of the auxiliary agent is added into 1/2MS culture medium-1NAA and 0.35-0.5 mg.L-1After gibberellin is generated, rootless seedlings can root; finally, 0.25-0.3 mg.L of the culture medium is added into the MS culture medium-1NAA is beneficial to the culture of rooting seedlings and strong seedlings. In addition, there is a report on the use of seeds and anthers to develop tissue culture seedlings.
There are also many reports on the researches on the detoxification of the passion fruit seedlings. Zhouyoulin (CN 1O 6258965A) discloses a cultivation method of passion fruit virus-free seedlings, wherein the main virus-free mode is that stem tips are sterilized by hot water and then are pre-cultured to induce adventitious buds, the obtained tissue culture seedlings are subjected to heat treatment for 3-6 days at 40-45 ℃, and stem tips with 0.2cm length of new axillary buds are stripped to cultivate virus-free seedlings; meanwhile, a plurality of antibacterial agents with different proportions are added into the culture medium. Wenzhidine (CN 1O625897 OA) discloses a passion fruit virus-free seedling cultivation method comprising the technical links of seed heat treatment, seed disinfection treatment, stock culture, stem tip disinfection treatment, stem tip heat treatment, scion propagation, stem tip grafting, rooting culture, detection and the like. Chinese patent application publication numbers CN105284314A, CN105706935A and the like all disclose cultivation methods of passion fruit virus-free seedlings, and CN105519418A also discloses a transplantation method of passion flower tissue culture seedlings. The existing passion flower seedling detoxification methods have the problems that the operation procedures are complex, explants are derived from mature organs or tissues which may carry viruses and germs, the heat treatment detoxification effect of a single stem tip is unstable, and the like. The newly germinated top shoot of passion flower belongs to tender tissue with strong differentiation capability, almost has no virus or germ, and is easy to brown after disinfection treatment, so the tissue culture seedling raising method for inducing axillary bud differentiation, callus differentiation and cluster bud differentiation by using the combination of cytokinin TDZ (thidiazuron) and auxin NAA as an explant and adopting the combination of the cytokinin TDZ and the auxin NAA is not reported at present.
Disclosure of Invention
The invention aims to provide a tissue culture seedling method which can obviously reduce the browning rate and the pollution rate of young tips of passion fruit canna and improve the initial induction survival rate, the cluster bud differentiation rate, the transplanting survival rate and the like of the tips of the passion fruit canna, and provides a reference basis for the industrial seedling utilization of improved seeds of the passion fruit canna.
The tissue culture seedling raising method for the passion fruit top shoots provided by the inventor comprises the following steps:
(1) explant disinfection: collecting immature tops of newly germinated passion fruit leaves of 3-5 days as explants; after washing, 4500mg.L-1Vitamin C +1000 mg.L-1Soaking in distilled water solution of citric acid for 45min, and adding 2500mg.L-1Soaking in distilled water solution of potassium permanganate for 30min, washing with distilled water for 3-5 times, and adding 1000 mg.L-1Sterilizing the mercuric chloride aqueous solution for 6min by conventional method, and washing with distilled water for 5 times;
(2) determination of the minimal medium: the basic culture medium of each stage of plant cell tissue culture is as follows: 1/2MS +12 g.L-1Agar +30 g.L-1Sucrose, pH 5.5; the culture conditions at each stage are as follows: the temperature is 24-26 ℃, the illumination intensity is 1500-2000 Lux, and the illumination time is 12-14 h/d;
(3) primary induction culture of axillary buds: vertically inoculating the sterilized explant according to the growth polarity, and inducing axillary buds to sprout new leaves; after culturing for 20 days, germinating 2-3 new leaves; the formula of the culture medium is as follows: minimal medium+1.2g.L-1AC+1.2 g.L-1PVP+0.5mg.L-1 TDZ+0.05 mg.L-1NAA；
(4) Callus culture: cutting the leaf blade into 1cm2Inoculating after the small blocks, and inducing callus differentiation; after culturing for 35d, green loose callus with the diameter of 1.6cm is generated; the formula of the culture medium is as follows: minimal medium +1.2g.L-1AC+1.2g.L-1 PVP +1.2 mg.L-1 TDZ+0.1mg.L-1NAA；
(5) Subculture multiplication culture: gently dividing the callus into small pieces with diameter of 0.5cm by using forceps, and inoculating to induce cluster bud differentiation; after culturing for 40 days, 5-7 cluster buds are differentiated from the callus. The formula of the culture medium is as follows: minimal medium +1.2g.L-1AC+1.2g.L-1 PVP +1.5 mg.L-1 TDZ +0.03mg.L-1NAA。
(6) Rooting induction culture: cutting single buds of cluster buds formed by subculture proliferation, inoculating the cluster buds to a rooting culture medium, inducing to root, culturing for 35 days, wherein 90% of the cluster buds generate root systems, 5-10 roots of each plant are obtained, and the root length is 0.5-3.0 cm, and the formula of the culture medium is as follows: minimal medium +0.1mg.L-1 6-BA+1.2 mg.L-1 IBA。
(7) Transplanting bottle seedlings: culturing the roots for 35 days, opening a bottle cap of a bottle seedling with the height of more than 8cm, adding 20ml of distilled water into each bottle, and hardening and culturing the seedlings in a culture room at the temperature of 24-26 ℃ for 10 days; transplanting seedlings after preparing a matrix; meanwhile, the seedbed is shaded, and the survival rate is more than 95 percent after 1 month of transplanting.
In the step (7), the seedling exercising method comprises the following steps: culturing for 35 days, opening the bottle cover of the bottle seedling with the height of more than 8cm, adding 20ml of distilled water into each bottle, and exercising the seedling for 10 days in a culture room at the temperature of 24-26 ℃; the matrix is prepared by mixing perlite and sandy yellow soil at volume ratio of 1: 2.5, packaging into 5cm × 5cm nutrition bag, and adding into 3500 mg.L-1Sterilizing the carbendazim aqueous solution to obtain the product; the seedling transplanting is to take out the bottle seedlings after hardening, clean the root system with water, place the bottle seedlings in a magnetic disk with a small amount of water, implant the seedlings in a sterilized and bagged matrix, and plant 1 in each bag; the seedbed is used for shading the seedbed by placing the seedling planting nutrition bags one by onePutting the seeds into a mud-bottom seedbed with the width of 1m and the depth of 0.3cm, and then watering the seeds thoroughly; a small arched shed with the height of 0.6m is built on the seedbed, and a white mulching film and a sunshade net with the sunshade rate of 40% are covered on the arched shed.
The inventor points out that: in the steps (4), (5) and (6) of the method, 1.2g.L of the culture medium is added-1AC and 1.2g.L-1PVP is used for adsorbing harmful substances secreted in the passion flower tissue culture process and inhibiting generation of phenolic browning substances.
The innovation points of the invention are as follows: according to the passion fruit passion flower apical bud tissue culture seedling culture method provided by the invention, firstly, antioxidant Vc and citric acid distilled water solution are adopted for soaking, so that the antioxidant and browning resistant capabilities of young and tender explants are obviously improved; then, potassium permanganate distilled water solution is adopted for soaking sterilization, 75% ethanol is not used for sterilization, and the activity of the explant is obviously improved; finally, 1.2g.L of the culture medium is added into the culture medium of the three stages of primary induction, callus induction and subculture proliferation-1AC and 1.2g.L-1PVP is used for adsorbing harmful substances secreted in the passion flower tissue culture process and inhibiting generation of browning substances such as phenols. After the sterilization and anti-browning treatment method is adopted, the initial induction pollution rate and the browning rate can be controlled to be below 10%, and the multiplication coefficient is above 5.0. The original technology is that the tender top shoot of passion fruit is used as an explant, and the combination of cytokinin TDZ and auxin NAA is adopted to realize the tissue culture seedling method of passion fruit.
Detailed Description
Example 1: test of 2016 at 4 months
(1) Explant disinfection: collecting 50 passionflower spring shoots of the newly germinated 3d purple fruits, washing with tap water, washing with distilled water for 3 times, and shearing top young shoots with the length of 3-4 cm as explants; using 4500
mg.L-1Vc+1000mg.L-1Soaking in distilled citric acid solution for 45min, and washing with distilled water for 5 times. On a clean bench, 2500mg.L-1Soaking in potassium permanganate distilled water solution for 30min, and washing with distilled water for 3 times; cutting the explant into 2.0cm long top shoots, and adding 1000 mg.L-1Sterilizing with mercuric chloride water solution for 6min, and washing with distilled water for 5 times;
(2) primary induction culture of axillary buds: vertically inoculating the sterilized explants according to growth polarity, wherein the formula of a culture medium is as follows: minimal medium +1.2g.L-1AC+1.2 g.L-1 PVP +0.5mg.L-1TDZ +0.05mg.L-1 50 NAAs are inoculated, after 10 days of culture, 4 are dead in brown stain, and the brown stain rate is 8.0 percent; after the rest of 20 days, 2-3 new leaves are developed, and the initial induction survival rate is 92.0%;
(3) callus induction culture: when the area of axillary bud new leaf reaches 2.0cm2Dividing the new leaf 1 into two parts, and cutting into 1.0cm2Inoculating after the small pieces, and preparing a culture medium formula: minimal medium +1.2g.L-1AC +1.2g.L-1 PVP+1.2 mg.L-1TDZ+0.1mg.L-1NAA. Inoculating 80 pieces of the callus, culturing for 35 days, and generating 73 green loose callus tissues with the diameter of 1.6 cm; 7 browned deaths; the callus induced differentiation rate is 91.3%;
(4) subculture multiplication culture: gently dividing the callus into small pieces with diameter of 0.5cm by using forceps, and then inoculating, wherein the culture medium formula is as follows: minimal medium +1.2g.L-1AC+1.2g.L-1 PVP +1.5mg.L-1 TDZ+0.03mg.L-1NAA. 200 calluses are inoculated in total, 5-7 cluster buds are differentiated from 184 calluses after the calluses are cultured for 40 days, and the multiplication coefficient is 5.1;
(5) rooting induction culture: after subculture for 50 days, the average length of the cluster buds reaches more than 1.5cm, the cluster buds are cut into single buds and inoculated, and the culture medium formula is as follows: minimal medium +0.1mg.L-1 6-BA+1.2 mg.L-1The IBA is inoculated into 800 plants in total, 723 roots germinate after 35 days of culture, 5-10 roots of each plant grow, and the rooting rate is 90.4%;
(6) the minimal medium at each stage is: 1/2MS +12 g.L-1Agar +30 g.L-1Sucrose, pH 5.5; the culture conditions at each stage are as follows: the temperature is 24-26 ℃, the illumination is 1500-2000 Lux, and the illumination time is 12-14 h/d.
(7) Transplanting bottle seedlings: carrying out rooting culture for 35d, uncovering a bottle cap of a bottle seedling with the height of more than 8cm, adding 20ml of distilled water into each bottle, and carrying out seedling hardening culture for 10d in a culture room at the temperature of 24-26 ℃; placing the perlite and sandy yellow soil mixed matrix with the volume ratio of 1: 2.5 into a nutrition bag of 5cm multiplied by 5cm, and adding 3500 mg.L-1The aqueous solution of carbendazim is sterilized; taking out the bottle seedlings after hardening, cleaning root systems with water, placing the bottle seedlings in a magnetic disk with a small amount of water, and implanting the bottle seedlings into a sterilized and bagged matrix, wherein 1 plant is planted in each bag; placing the seedling planting nutrition bags into a mud bottom seedbed with the width of 1m and the depth of 0.3cm, and then watering thoroughly; setting up a 0.6m arched shed on the seedbed, covering with white ground film and sunshade net with shading rate of 40%, transplanting 600 plants together, examining after 1 month, and transplanting to obtain 572 plants which survive with survival rate of 95.3%.
Example 22017 year 4 month test
(1) Explant disinfection: collecting 50 spring shoots of Passiflora incarnata which just germinates for 5 days, washing with tap water, washing with distilled water for 3 times, cutting top shoots with length of 3-4 cm as explant, and adding 4500mg.L of the explant-1Vc+1000mg.L-1Soaking in distilled water solution of citric acid for 45min, washing with distilled water for 5 times, and treating with 2500mg.L on a clean bench-1Soaking in distilled water solution of potassium permanganate for 30min, washing with distilled water for 3 times, cutting the explant into 2.0cm tip, and adding 1000 mg.L-1Sterilizing with mercuric chloride water solution for 6min, and washing with distilled water for 5 times.
(2) Primary induction culture of axillary buds: vertically inoculating the sterilized explants according to growth polarity, wherein the formula of a culture medium is as follows: minimal medium +1.2g.L-1AC+1.2 g.L-1 PVP +0.5mg.L-1TDZ +0.05mg.L-1 NAA. Inoculating 50 seeds in total, culturing for 10 days, and then dying 2 seeds with browning rate of 4.0%; and after the rest of 20 days, 2-3 new leaves are developed, and the initial induction survival rate is 92.0%.
(3) Callus induction culture: when the area of axillary bud new leaf reaches 2.0cm2Dividing the new leaf 1 into two parts, and cutting into 1.0cm2Inoculating after the small pieces, and preparing a culture medium formula: minimal medium +1.2g.L-1AC +1.2g.L-1 PVP +1.2 mg.L-1TDZ+0.1mg.L-1NAA. Inoculating 80 pieces of the above-mentioned materials, culturing for 35d, 75 green loose callus whose diameter is 1.6cm can be produced; 5 browning deaths; the callus induced differentiation rate is 93.8%.
(4) Subculture multiplication culture: gently dividing the callus into small pieces with diameter of 0.5cm by using forceps, and then inoculating, wherein the culture medium formula is as follows: minimal medium+1.2g.L-1AC+1.2g.L-1 PVP+1.5 mg.L-1 TDZ +0.03mg.L-1NAA. 200 are inoculated in total, 5-7 cluster buds are differentiated from 189 callus tissues after culture for 40 days, and the multiplication coefficient is 5.3.
(5) Rooting induction culture: after subculture for 50 days, the average length of the cluster buds reaches more than 1.5cm, the cluster buds are cut into single buds and inoculated, and the culture medium formula is as follows: minimal medium +0.1mg.L-1 6-BA+1.2 mg.L-1IBA. And (3) inoculating 800 seeds in total, and culturing for 35 days to obtain 736 germination roots, wherein 5-10 roots are planted in each plant, and the rooting rate is 92.0%.
(6) The basic culture medium of each stage of plant cell tissue culture is as follows: 1/2MS +12 g.L-1Agar +30 g.L-1Sucrose, pH 5.5; the culture conditions at each stage are as follows: the temperature is 24-26 ℃, the illumination is 1500-2000 Lux, and the illumination time is 12-14 h/d.
Claims (2)
1. A tissue culture seedling raising method for passion fruit top shoots is characterized by comprising the following steps:
(1) explant disinfection: collecting immature tops of newly germinated passion fruit leaves of 3-5 days as explants; after washing, 4500mg.L is used-1Vitamin C +1000 mg. L-1Soaking citric acid distilled water solution for 45min, and soaking with 2500mg.L-1Soaking in distilled water solution of potassium permanganate for 30min, washing with distilled water for 3-5 times, and adding 1000 mg.L-1Sterilizing the mercuric chloride aqueous solution for 6min by conventional method, and washing with distilled water for 5 times;
(2) determination of the minimal medium: the basic culture medium of each stage of plant cell tissue culture is as follows: 1/2MS +12 g.L-1Agar +30 g. L-1Sucrose, pH 5.5; the culture conditions at each stage are as follows: the temperature is 24-26 ℃, the illumination intensity is 1500-2000 Lux, and the illumination time is 12-14 h/d;
(3) primary induction culture of axillary buds: vertically inoculating the sterilized explant according to the growth polarity, and inducing axillary buds to sprout new leaves; after culturing for 20 days, germinating 2-3 new leaves; the formula of the culture medium is as follows: minimal medium +1.2g.L-1Activated carbon +1.2g.L-1Polyvinylpyrrolidone +0.5mg· L-1Thidiazuron +0.05 mg. L-1Naphthylacetic acid;
(4) callus culture: cutting the leaf blade into 1cm2Inoculating after the small blocks, and inducing callus differentiation; after culturing for 35d, green loose callus with the diameter of 1.6cm is generated; the formula of the culture medium is as follows: minimal medium +1.2g.L-1Activated carbon +1.2g.L-1Polyvinylpyrrolidone +1.2 mg. L-1Thidiazuron +0.1mg. L-1Naphthylacetic acid;
(5) subculture multiplication culture: gently dividing the callus into small pieces with diameter of 0.5cm by using forceps, and inoculating to induce cluster bud differentiation; after culturing for 40 days, differentiating 5-7 cluster buds from the callus; the formula of the culture medium is as follows: minimal medium +1.2g.L-1Activated carbon +1.2g.L-1Polyvinylpyrrolidone +1.5 mg. L-1Thidiazuron +0.03 mg. L-1Naphthylacetic acid;
(6) rooting induction culture: cutting single buds of cluster buds formed by subculture proliferation, inoculating the cluster buds to a rooting culture medium, inducing to root, culturing for 35 days, wherein 90% of the cluster buds generate root systems, 5-10 roots of each plant are obtained, and the root length is 0.5-3.0 cm, and the formula of the culture medium is as follows: minimal medium +0.1mg. L-16-benzylaminopurine +1.2 mg. L-1Indolebutyric acid;
(7) transplanting bottle seedlings: carrying out rooting culture for 35d, uncovering a bottle cap of a bottle seedling with the height of more than 8cm, adding 20ml of distilled water into each bottle, and carrying out seedling hardening culture for 10d in a culture room at the temperature of 24-26 ℃; transplanting seedlings after preparing a matrix; meanwhile, seedbed shading is carried out, and after the seedlings are transplanted for 1 month, the survival rate is over 95 percent.
2. The tissue culture seedling raising method for the passion fruit top shoot as claimed in claim 1, characterized in that in the step (7), the seedling exercising method comprises the following steps: carrying out rooting culture for 35d, uncovering a bottle cap of a bottle seedling with the height of more than 8cm, adding 20ml of distilled water into each bottle, and carrying out seedling hardening culture for 10d in a culture room at the temperature of 24-26 ℃; the matrix is prepared by mixing perlite and sandy yellow soil at volume ratio of 1: 2.5, packaging into 5cm × 5cm nutrition bag, and adding into 3500 mg.L-1Sterilizing the carbendazim aqueous solution to obtain the product; transplanting the seedlingsTaking out the bottle seedlings after hardening, cleaning root systems with water, placing the bottle seedlings in a porcelain dish with a small amount of water, and implanting the bottle seedlings into a sterilized and bagged matrix, wherein 1 plant is planted in each bag; the seedbed shading is to place the seedling planting nutrition bags into a muddy bottom seedbed with the width of 1m and the depth of 0.3cm in a row, and then to pour water; a small shed with the height of 0.6m is erected on the seedbed, and a white mulching film and a sunshade net with the shading rate of 40 percent are covered on the shed.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104106465A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
-
2018
- 2018-03-13 CN CN201810204586.1A patent/CN108419675B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104106465A (en) * | 2014-06-12 | 2014-10-22 | 南京工业大学大丰海洋产业研究院 | Explant disinfection method in plant tissue culture |
Non-Patent Citations (6)
| Title |
|---|
| Effect of plant growth regulators and antioxidants on in vitro plant regeneration and callus induction from leaf explants of purple passion fruit (Passiflora edulis Sims);Yoon Sun Huh et al.;《J Plant Biotechnol》;20171231(第44期);摘要和材料与方法部分、表1-4及图1 * |
| In vitro plant regeneration of Passiflora setacea D.C.(Passifloraceae): the influence of explant type, growth regulators, and incubation conditions;Lorena Melo Vieira et al.;《In Vitro Cell.Dev.Biol.—plant》;20141231(第50期);第738-745页 * |
| 植物生长调节剂对紫果西番莲离体培养的影响;陈发兴;《福建果树》;20020331(第121期);第7-8页 * |
| 百香果组织培养及植株再生;杨冬业等;《北方园艺》;20121231(第3期);第125-127页 * |
| 西番莲愈伤组织诱导研究;丰锋等;《西南农业大学学报（自然科学版）》;20061031;第28卷(第5期);第802-804页 * |
| 西番莲离体快繁研究;董丽兰等;《安徽农业科学》;20161231;第44卷(第24期);第160-162页 * |
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