CN113100062A - Method for obtaining haploid by culturing amorphophallus konjac ovule - Google Patents
Method for obtaining haploid by culturing amorphophallus konjac ovule Download PDFInfo
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- CN113100062A CN113100062A CN202110459769.XA CN202110459769A CN113100062A CN 113100062 A CN113100062 A CN 113100062A CN 202110459769 A CN202110459769 A CN 202110459769A CN 113100062 A CN113100062 A CN 113100062A
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- 241001312219 Amorphophallus konjac Species 0.000 title claims abstract description 30
- 235000001206 Amorphophallus rivieri Nutrition 0.000 title claims abstract description 30
- 238000000034 method Methods 0.000 title claims abstract description 21
- 238000012258 culturing Methods 0.000 title claims abstract description 20
- 210000001672 ovary Anatomy 0.000 claims abstract description 52
- 206010020649 Hyperkeratosis Diseases 0.000 claims abstract description 48
- 230000004069 differentiation Effects 0.000 claims abstract description 28
- 239000001963 growth medium Substances 0.000 claims abstract description 24
- 230000006698 induction Effects 0.000 claims abstract description 20
- 235000010485 konjac Nutrition 0.000 claims abstract description 18
- 241000196324 Embryophyta Species 0.000 claims abstract description 16
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 5
- 238000005286 illumination Methods 0.000 claims description 18
- 239000002609 medium Substances 0.000 claims description 16
- 239000005720 sucrose Substances 0.000 claims description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 9
- 239000008272 agar Substances 0.000 claims description 9
- 239000005556 hormone Substances 0.000 claims description 7
- 229940088597 hormone Drugs 0.000 claims description 7
- 229920001817 Agar Polymers 0.000 claims description 6
- 239000005708 Sodium hypochlorite Substances 0.000 claims description 6
- 239000007640 basal medium Substances 0.000 claims description 6
- 238000002791 soaking Methods 0.000 claims description 6
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 claims description 6
- 238000005406 washing Methods 0.000 claims description 6
- 239000003357 wound healing promoting agent Substances 0.000 claims description 5
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 4
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 3
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- 208000027418 Wounds and injury Diseases 0.000 claims description 3
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- 239000000843 powder Substances 0.000 claims description 3
- 239000008223 sterile water Substances 0.000 claims description 3
- 239000008399 tap water Substances 0.000 claims description 3
- 235000020679 tap water Nutrition 0.000 claims description 3
- QGHREAKMXXNCOA-UHFFFAOYSA-N thiophanate-methyl Chemical compound COC(=O)NC(=S)NC1=CC=CC=C1NC(=S)NC(=O)OC QGHREAKMXXNCOA-UHFFFAOYSA-N 0.000 claims description 3
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a method for obtaining haploid by culturing konjak ovule, which is characterized by comprising the following steps of S1: obtaining a female flower section of the non-pollinated konjak from a konjak flower plant; s2: carrying out low-temperature pretreatment and explant disinfection on female flower sections of amorphophallus konjac; s3: stripping off the ovary from the sterilized pedicel of the female flower section, and performing pre-culture; s4: dissecting the ovary under aseptic condition after the ovary is enlarged and the ovary wall is cracked and part of the ovary has callus, taking out ovules, and carrying out ovule callus induction culture; s5: when the diameter of the callus is more than 2cm, inoculating the induced callus into a differentiation culture medium, and carrying out differentiation culture on the ovule callus; s6: and (4) carrying out rooting culture on the bud obtained by differentiating the ovule callus to obtain a complete haploid plant. The haploid is obtained by culturing the non-pollinated ovary ovules of amorphophallus konjac, a set of amorphophallus konjac ovule culture regeneration system is established, and a new technical platform is provided for amorphophallus konjac haploid breeding.
Description
Technical Field
The invention relates to the technical field of plant tissue culture, in particular to a method for obtaining haploid by culturing amorphophallus konjac ovule.
Background
Konjak is a perennial herb of the family Araceae, contains a large amount of soluble dietary fiber glucomannan, is the seventh nutrient of a human body, is ten health-care foods determined by the United nations, and has obvious effects of reducing three highs, losing weight, relaxing the bowels and the like. Konjak is mainly distributed in places such as Sichuan, Yunnan, Shaanxi, Ningxia and Gansu in China, and the flower konjak is most widely distributed as a main cultivated species and is distributed in all mountain areas and hilly lands in the south of Qinling mountains.
Konjak is highly heterozygous and has a complicated genetic background, so that it is difficult to obtain a homozygous inbred line by a conventional method. Haploid is induced by anther or unpolarized ovule culture, pure line can be obtained after doubling, breeding period can be shortened, and breeding efficiency can be improved. In addition, the haploid can express recessive characters, can quickly screen useful genotypes, and improves the selection efficiency. However, no relevant research reports are found in culturing by using konjak anthers or unpolished ovules, so that the application of konjak haploid breeding is greatly limited.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a method for obtaining haploid by culturing amorphophallus konjac ovules, which utilizes the non-pollinated ovary ovules of amorphophallus konjac to obtain the haploid, establishes a set of amorphophallus konjac ovule culturing and regenerating system and provides a new technical platform for breeding the haploid of the amorphophallus konjac.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for culturing and obtaining haploid by utilizing an ovule of amorphophallus konjac is characterized by comprising the following steps,
s1: obtaining a female flower section of the non-pollinated konjak from a konjak flower plant;
s2: carrying out low-temperature pretreatment and explant disinfection on female flower sections of amorphophallus konjac;
s3: stripping off the ovary from the sterilized pedicel of the female flower section, and performing pre-culture;
s4: dissecting the ovary under aseptic condition after the ovary is enlarged and the ovary wall is cracked and part of the ovary has callus, taking out ovules, and carrying out ovule callus induction culture;
s5: when the diameter of the callus is more than 2cm, inoculating the induced callus into a differentiation culture medium, and carrying out differentiation culture on the ovule callus;
s6: and (4) carrying out rooting culture on the bud obtained by differentiating the ovule callus to obtain a complete haploid plant.
Further, the specific operation of step S1 includes the following steps,
s101: after the flower konjak spathe is opened, the accessory emits rotten dead body flavor, the anther does not loose powder, the scissors are sterilized by 75% of alcohol, and the male flower section and the spathe are cut off;
s102: smearing a plant wound healing agent on the wound, reserving female flower sections, and bagging to prevent insect-borne pollination;
s103: and when the ovary on the female flower is changed from white to light green, cutting off the whole female flower section for later use.
Further, the plant wound healing agent in step S102 includes thiophanate-methyl.
Further, the specific operation of step S2 includes the following steps,
s201: pre-treating female flower sections of amorphophallus konjac in a refrigerator at 4 ℃ for 2 days;
s202: washing the whole female flower section in tap water for 10-20min, and soaking in 75% alcohol for 30-50 s; the female flower section comprises a pedicel and an ovary;
s203: soaking the female flower section soaked in alcohol in sodium hypochlorite for 8-10 min;
s204: and (3) washing the female flower sections soaked and disinfected by sodium hypochlorite for 3-5 times by using sterile water, and then sucking dry by using sterile filter paper for later use.
Further, the specific operation of step S3 includes the following steps,
s301: stripping off the ovary from the sterilized pedicel of the female flower section;
s302: cutting off the stigma part on the ovary, inoculating the rest part into MS basal medium without hormone addition, and culturing for 10-15 days;
the MS basal medium contains 35g/L of sucrose and 5g/L of agar;
the culture conditions were: the illumination intensity is 2000-30001ux, the illumination time is 16h/d, and the temperature is 28 +/-1 ℃.
Further, the specific operation of step S4 includes the following steps,
s401: dissecting the ovary under aseptic condition after the ovary expands and the ovary wall changes from light green to dark green and cracks and part of the ovary wall has callus, removing the ovary wall, and taking out ovules;
s402: inoculating the ovule into a callus induction culture medium for callus induction culture;
the callus induction culture medium takes MS as a basic culture medium, and 0-3 mg.L is added-10 to 1.5 mg.L of NAA-1Of 6-BA, 35 g.L-1Sucrose of (5 g. L)-1And the callus induction medium has a pH of 5.8;
the culture conditions comprise that the culture is performed in a dark environment for 10-15 days, then the illumination culture is performed, the illumination intensity is 2000 and 30001ux, the illumination time is 14h/d, and the temperature is 26 +/-1 ℃.
Further, the differentiation medium described in step S5 is MS minimal medium, and is added at a concentration of 0-1.5 mg.L-1The NAA of (1) is 0-3.0 mg.L-16-BA, 30 g.L-1Sucrose of (3), 7 g.L-1The agar of (4); and the differentiation medium has a pH of 5.8;
the culture conditions for the differentiation culture of the ovule callus are as follows: the illumination intensity is 4000-.
Further, the differentiation medium described in step S5 was MS minimal medium added at a concentration of 1.0 mg.L-1NAA of (1.0 mg. L)-16-BA of (1).
Further, the specific operation of step S6 includes: transferring the bud obtained by differentiation into 1/2MS culture medium, adding the bud with the concentration of 0-1.5 mg.L-1The NAA of (1) is 0-3.0 mg.L-16-BA, 30 g.L-1Sucrose, 5 g.L-1Agar, 2 g.L-1Activated carbon, culturing adventitious bud to obtain complete haploid plant, and MS culturingThe pH of the base was 5.8.
Further, the concentration of NAA added into the culture medium for adventitious bud rooting culture is 0.5 mg.L-1The concentration of 6-BA is 2.0 mg.L-1。
The invention has the beneficial effects that:
1. according to the method, the haploid is obtained by culturing the amorphophallus konjac ovule, the haploid is obtained by culturing the non-pollinated ovary ovule of the amorphophallus konjac, a set of amorphophallus konjac ovule culture regeneration system is established, and a new technical platform is provided for breeding the haploid of the amorphophallus konjac;
2. according to the invention, the female flower sections are subjected to double disinfection treatment by using different disinfectants to obtain sterile explants, so that the pollution in the culture process is reduced;
3. because the ovule of the amorphophallus konjac is extremely small, the ovule in the ovary can be prevented from being damaged by pre-culturing the unpolished ovary;
4. according to the method, the female flower segments are pretreated at low temperature, and the proportion of the culture medium is screened at different culture stages, so that the induction rate of the callus is effectively improved.
Drawings
FIG. 1 shows callus induction in the present invention;
FIG. 2 is the differentiation culture of callus according to the present invention;
FIG. 3 shows the haploid state obtained in the present invention.
Detailed Description
In order to make those skilled in the art better understand the technical solution of the present invention, the following further describes the technical solution of the present invention with reference to the drawings and the embodiments.
A method for culturing amorphophallus konjac ovule to obtain haploid comprises the following steps,
s1: obtaining a female flower section of the non-pollinated konjak from a konjak flower plant;
specifically, S101: after the flower konjak spathe is opened, the accessory emits rotten dead body flavor, the anther does not loose powder, the scissors are sterilized by 75% of alcohol, and the male flower section and the spathe are cut off;
s102: smearing plant wound healing agent thiophanate-methyl on the wound, reserving female flower segments, and bagging to prevent insect-borne pollination;
s103: and when the ovary on the female flower is changed from white to light green, cutting off the whole female flower section for later use.
Further, step S2: carrying out low-temperature pretreatment and explant disinfection on female flower sections of amorphophallus konjac;
specifically, S201: pre-treating female flower sections of amorphophallus konjac in a refrigerator at 4 ℃ for 2 days;
s202: washing the whole female flower section in tap water for 10-20min, and soaking in 75% alcohol for 30-50 s; the female flower section comprises a pedicel and an ovary;
s203: soaking the female flower section soaked in alcohol in sodium hypochlorite for 8-10 min;
s204: and (3) washing the female flower sections soaked and disinfected by sodium hypochlorite for 3-5 times by using sterile water, and then sucking dry by using sterile filter paper for later use.
Further, step S3: stripping off the ovary from the sterilized pedicel of the female flower section, and performing pre-culture;
specifically, S301: stripping off the ovary from the sterilized pedicel of the female flower section;
s302: cutting off the stigma part on the ovary, inoculating the rest part into MS basal medium without hormone addition, and culturing for 10-15 days;
the MS basal medium contains 35g/L of sucrose and 5g/L of agar;
the culture conditions were: the illumination intensity is 2000-30001ux, the illumination time is 16h/d, and the temperature is 28 +/-1 ℃.
Further, step S4: dissecting the ovary under aseptic condition after the ovary is enlarged and the ovary wall is cracked and part of the ovary has callus, taking out ovules, and carrying out ovule callus induction culture;
specifically, S401: dissecting the ovary under aseptic condition after the ovary expands and the ovary wall changes from light green to dark green and cracks and part of the ovary wall has callus, removing the ovary wall, and taking out ovules;
s402: the ovule is inoculated into callus induction culture medium for callus induction culture, as shown in figure 1.
The callus induction culture medium takes MS as a basic culture medium, and 0-0.3 mg.L is added-10 to 1.5 mg.L of NAA-10 to 1.5 mg.L of 6-BA-135 g.L of hormone (D)-1Sucrose of (5 g. L)-1And the callus induction medium has a pH of 5.8;
the culture conditions comprise that the culture is performed in a dark environment for 10-15 days, then the illumination culture is performed, the illumination intensity is 2000 and 30001ux, the illumination time is 14h/d, and the temperature is 26 +/-1 ℃.
Preferably, the addition amount of NAA in the callus induction culture medium is 1.0 mg.L-1The addition amount of 6-BA is 1.0 mg.L-1。
Further, step S5: when the callus expands to a diameter of 2cm or more, inoculating the induced callus into a differentiation medium, and performing differentiation culture of the ovule callus as shown in figure 2.
Specifically, the differentiation culture medium takes MS as a basic culture medium, and the concentration of the MS is 0-1.5 mg.L-1The NAA of (1) is 0-3.0 mg.L-16-BA in a concentration of 0 to 3.0 mg.L-130 g.L of hormone (D)-1Sucrose of (3), 7 g.L-1The agar of (4); and the differentiation medium has a pH of 5.8;
the culture conditions for the differentiation culture of the ovule callus are as follows: the illumination intensity is 4000-.
Furthermore, the concentration of NAA and 6-BA in the differentiation medium is optimized in the invention, and the results of comparative experiments are shown in the following table 1 by adopting different NAA and 6-BA concentrations.
TABLE 1 Effect of different hormone concentration ratios on callus differentiation
As can be seen from Table 1, when the concentration of 6-BA was 1.0 mg.L-1The concentration of NAA is 1.0 mg.L-1The callus induction rate is the highest and reaches 90%, and the weight gain of the inoculation block is the largest and reaches 4.6 times.
Further, step S6: rooting culture is carried out on the bud obtained by differentiation of the ovule callus, and a complete haploid plant is obtained, as shown in figure 3.
Specifically, the bud obtained by differentiation is transferred into 1/2MS culture medium, and the concentration is 0-1.5 mg.L-1The NAA of (1) is 0-3.0 mg.L-16-BA, 30 g.L-1Sucrose, 5 g.L-1Agar, 2 g.L-1And (3) carrying out adventitious bud rooting culture by using activated carbon to obtain a complete haploid plant, wherein the pH value of the MS culture medium is 5.8.
Furthermore, the invention optimizes the formula of the culture medium for adventitious bud rooting culture, adopts different NAA and 6-BA concentrations to carry out comparative experiments, and the results are shown in the following table 2.
TABLE 2 Effect of different hormone concentration ratios on adventitious bud differentiation
As can be seen from Table 2, when the concentration of 6-BA was 2.0 mg.L-1The concentration of NAA is 0.5 mg.L-1In the meantime, the adventitious bud differentiation rate is the highest and reaches 90%, and the inoculation increment coefficient is also the highest and is 4.6.
The adventitious bud rooting condition of amorphophallus konjac when different adventitious bud rooting media and NAA concentrations were selected is shown in Table 3 below.
TABLE 3 Effect of different media and NAA concentrations on rooting of adventitious buds of amorphophallus konjac
As can also be seen from Table 3, when the concentration of NAA was 0.5 mg.L-1In the process, the number of roots of the adventitious buds of the amorphophallus konjac is the largest, the number of the roots reaches as many as 21, the root length is the longest, and the average root length is more than 9 cm.
The foregoing shows and describes the general principles, essential features, and advantages of the invention. It will be understood by those skilled in the art that the present invention is not limited to the embodiments described above, which are described in the specification and illustrated only to illustrate the principle of the present invention, but that various changes and modifications may be made therein without departing from the spirit and scope of the present invention, which fall within the scope of the invention as claimed. The scope of the invention is defined by the appended claims and equivalents thereof.
Claims (10)
1. A method for culturing and obtaining haploid by utilizing an ovule of amorphophallus konjac is characterized by comprising the following steps,
s1: obtaining a female flower section of the non-pollinated konjak from a konjak flower plant;
s2: carrying out low-temperature pretreatment and explant disinfection on female flower sections of amorphophallus konjac;
s3: stripping off the ovary from the sterilized pedicel of the female flower section, and performing pre-culture;
s4: dissecting the ovary under aseptic condition after the ovary is enlarged and the ovary wall is cracked and part of the ovary has callus, taking out ovules, and carrying out ovule callus induction culture;
s5: when the diameter of the callus is more than 2cm, inoculating the induced callus into a differentiation culture medium, and carrying out differentiation culture on the ovule callus;
s6: and (4) carrying out rooting culture on the bud obtained by differentiating the ovule callus to obtain a complete haploid plant.
2. The method of claim 1, wherein the step S1 comprises the following steps,
s101: after the flower konjak spathe is opened, the accessory emits rotten dead body flavor, the anther does not loose powder, the scissors are sterilized by 75% of alcohol, and the male flower section and the spathe are cut off;
s102: smearing a plant wound healing agent on the wound, reserving female flower sections, and bagging to prevent insect-borne pollination;
s103: and when the ovary on the female flower is changed from white to light green, cutting off the whole female flower section for later use.
3. The method of claim 2, wherein the wound healing agent of the plant in step S102 comprises thiophanate-methyl.
4. The method for obtaining haploid by culturing an ovule of amorphophallus konjac as claimed in claim 2, wherein the specific operation of step S2 includes the following steps,
s201: pre-treating female flower sections of amorphophallus konjac in a refrigerator at 4 ℃ for 2 days;
s202: washing the whole female flower section in tap water for 10-20min, and soaking in 75% alcohol for 30-50 s; the female flower section comprises a pedicel and an ovary;
s203: soaking the female flower section soaked in alcohol in sodium hypochlorite for 8-10 min;
s204: and (3) washing the female flower sections soaked and disinfected by sodium hypochlorite for 3-5 times by using sterile water, and then sucking dry by using sterile filter paper for later use.
5. The method of claim 4, wherein the step S3 comprises the following steps,
s301: stripping off the ovary from the sterilized pedicel of the female flower section;
s302: cutting off the stigma part on the ovary, inoculating the rest part into MS basal medium without hormone addition, and culturing for 10-15 days;
the MS basal medium contains 35g/L of sucrose and 5g/L of agar;
the culture conditions were: the illumination intensity is 2000-30001ux, the illumination time is 16h/d, and the temperature is 28 +/-1 ℃.
6. The method of claim 5, wherein the step S4 comprises the following steps,
s401: dissecting the ovary under aseptic condition after the ovary expands and the ovary wall changes from light green to dark green and cracks and part of the ovary wall has callus, removing the ovary wall, and taking out ovules;
s402: inoculating the ovule into a callus induction culture medium for callus induction culture;
the callus induction culture medium takes MS as a basic culture medium, and 0-3 mg.L is added-10 to 1.5 mg.L of NAA-1Of 6-BA, 35 g.L-1Sucrose of (5 g. L)-1And the callus induction medium has a pH of 5.8;
the culture conditions comprise that the culture is performed in a dark environment for 10-15 days, then the illumination culture is performed, the illumination intensity is 2000 and 30001ux, the illumination time is 14h/d, and the temperature is 26 +/-1 ℃.
7. The method of claim 6, wherein the differentiation medium of step S5 is MS minimal medium, and is added at a concentration of 0-1.5 mg-L-1The NAA of (1) is 0-3.0 mg.L-16-BA, 30 g.L-1Sucrose of (3), 7 g.L-1The agar of (4); and the differentiation medium has a pH of 5.8;
the culture conditions for the differentiation culture of the ovule callus are as follows: the illumination intensity is 4000-.
8. The method of claim 7, wherein the differentiation medium of step S5 is MS minimal medium, and is added at a concentration of 1.0 mg-L-1NAA of (1.0 mg. L)-16-BA of (1).
9. The method for obtaining haploid by culturing an ovule of amorphophallus konjac as claimed in claim 8, wherein the specific operation of step S6 includes: transferring the bud obtained by differentiation into 1/2MS culture medium, adding the bud with the concentration of 0-1.5 mg.L-1At a concentration of NAA of0～3.0mg·L-16-BA, 30 g.L-1Sucrose, 5 g.L-1Agar, 2 g.L-1And (3) carrying out adventitious bud rooting culture by using activated carbon to obtain a complete haploid plant, wherein the pH value of the MS culture medium is 5.8.
10. The method of claim 9, wherein the NAA concentration of the culture medium for rooting adventitious buds is 0.5 mg-L-1The concentration of 6-BA is 2.0 mg.L-1。
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| CN107211891A (en) * | 2017-05-27 | 2017-09-29 | 安康市农业科学研究所 | A kind of live fast breeding technique of konjaku callus |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN107211891A (en) * | 2017-05-27 | 2017-09-29 | 安康市农业科学研究所 | A kind of live fast breeding technique of konjaku callus |
Non-Patent Citations (9)
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| LINGLING ZHAO等: "Embryogenesis and plant regeneration from", 《AFRICAN JOURNAL OF BIOTECHNOLOGY 》 * |
| 孔凡伦等: "从白魔芋未授粉子房培养出单倍体植株(简报)", 《植物学通报》 * |
| 段龙飞等: "花魔芋实生种子组织培养研究初探", 《山西农业大学学报(自然科学版)》 * |
| 王自布等: "魔芋不同外植体组培快繁及其再生体系的研究", 《分子植物育种》 * |
| 蔡阳光等: "不同激素对杂交魔芋愈伤诱导及植株再生的影响", 《热带农业科学》 * |
| 覃剑锋等: "培养环境条件对花魔芋花粉萌发的影响", 《山西农业大学学报(自然科学版)》 * |
| 谢庆华等: "不同外植体诱导魔芋微球茎的比较研究", 《云南农业大学学报》 * |
| 陈国爱: "花魔芋根状茎组培配方优化研究", 《蔬菜》 * |
| 陈国爱: "魔芋外植体的消毒方法研究", 《陕西农业科学》 * |
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