CN105494088A - Disease-resistant variety breeding method of wild ganoderma lucidum - Google Patents
Disease-resistant variety breeding method of wild ganoderma lucidum Download PDFInfo
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- CN105494088A CN105494088A CN201510965063.5A CN201510965063A CN105494088A CN 105494088 A CN105494088 A CN 105494088A CN 201510965063 A CN201510965063 A CN 201510965063A CN 105494088 A CN105494088 A CN 105494088A
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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
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
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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
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Abstract
The invention relates to a disease-resistant variety breeding method of wild ganoderma lucidum. An original strain is a mycelium cultured after wild ganoderma lucidum vegetative mass cells induce calluses, the original strain is inoculated in a PDA culture medium containing trichoderma viride spores and cultured in a constant-temperature box which is 25 DEG C, strong fruiting bodies are selected for tissue separation, separated strains are subjected to laboratory-scale cultivation, two best strains are screened out, the two strains are cultured into mycelia in cultivation bottles respectively and irradiated by gamma-ray with the radiation intensity of 0.58-0.76 r/day for 10 days, the strains with higher mycelia concentration degree are selected for rejuvenated culture, different strains are separated and purified, and the optimal strain is selected according to mycelia growth conditions to serve as a new disease-resistant variety. The ganoderma lucidum variety bred through the method is high in trichoderma viride resistance, and the problems that the ganoderma lucidum yield is low and the quality of the ganoderma lucidum is poor because of trichoderma viride can be prevented.
Description
Technical field
The present invention relates to glossy ganoderma breeding field, particularly relate to the disease-resistant variety selection of a kind of wild red sesame.
Background technology
The lucid ganoderma stock culture of China mostly is the kind introduced by Japan, Korea S, through cultivation repeatedly for many years, degenerate serious, 1/3 of tame effective component of glossy ganoderma deficiency Wild ganoderma, the principle active component such as GL-B, triterpene constantly lacks, and disease resistance high-temperature resistance is weak, output and quality is caused to decline year by year.Therefore, originate in Wild ganoderma using China and carry out as mother's kind the pole when business that disease-resistant variety seed selection becomes glossy ganoderma breeding.
Summary of the invention
In order to solve the problems of the technologies described above, a kind of breeding method of the wild red sesame disease-resistant variety primary for China southern area of Jiangsu Province is provided, the invention provides following technical scheme:
A disease-resistant variety selection for wild red sesame, comprises the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelia as the bacterial classification that sets out;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, and cultivate in 25 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and carries out lab scale cultivation to isolated strains, filters out two bacterial strains that mycelial growth situation is best; Select the strong bacterial strain of resistance against diseases as variation raw material;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.58-0.76 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties according to mycelial growth situation.Adopt gamma-rays to irradiate and can bring out mycelia variation; improve protective enzyme active; mutagenesis obtains disease-resistant varieties; select 0.58-0.76 human relations/sky this compared with high radiation dose carry out long-term irradiation treatment be because radiation dose little; mycelia absorbed dose is little; be not enough to grow on mycelial growth the impact producing mutability, radiation dose is excessive, can cause destructive impact to mycelia, and breeding of new variety cannot be completed.
Further, select bright color in step S1, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame.Select excellent mother to plant to be conducive to the gene of high-quality to be extended to new varieties.
Further, to strain inoculation be set out in the PDA medium containing Trichoderma viride spore in step S2, and cultivate in the insulating box of 32-35 DEG C, the fruit body choosing robust growth carries out tissue separation, and lab scale cultivation is carried out to isolated strains, filter out two bacterial strains that mycelial growth situation is best.Glossy ganoderma disease is mainly divided into infectious disease and non-infectious disease, and Trichoderma viride is infectious disease, and temperature is too high is non-infectious disease, and the bacterial classification that simultaneously will set out is placed in two kinds of simultaneous environment of disease, can make the resistance against diseases of new varieties more comprehensively.And, temperature is higher, Trichoderma viride growth rate is faster, a more harsh disease environment can be provided for the bacterial classification that sets out, make new varieties resistance against diseases stronger, cultivate under red sesame is placed in high temperature meanwhile, new varieties can be made to have the ability of high temperature resistance concurrently, and at greenhouse effect, more significant contemporary society also seems very important.
Further, strain inoculation will be set out in the PDA medium containing Trichoderma viride spore, mould, Paecilomyces varioti and aspergillus spore in step S2.Glossy ganoderma infectious disease Green wood is mould is that occurrence frequency and the extent of injury are the highest, but mould, Paecilomyces varioti and aspergillus infection probability are also higher, therefore add this 4 miospore in the medium simultaneously and new varieties can be made to possess the resistivity of multiple infectious disease.
Further, in step S3, gamma-rays exposure intensity is 0.60-0.65 human relations/sky.Mycelia sudden change under this radiation dose is the most active, and the red sesame new varieties resistance against diseases obtained is strong.
Beneficial effect of the present invention is: the wild red sesame new varieties disease resistance ability adopting this method to cultivate is strong and comprehensively, can retain the merit of wild red sesame, output is high, active ingredient is high simultaneously.
Embodiment
Embodiment 1,
A disease-resistant variety selection for wild red sesame, comprises the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelia as the bacterial classification that sets out; Select bright color in the present embodiment, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, by this medium called after A1, and cultivate in 25 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and lab scale cultivation is carried out to isolated strains, filter out two bacterial strains that mycelial growth situation is best;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.62 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties SH-1 according to mycelial growth situation.
Embodiment 2,
A disease-resistant variety selection for wild red sesame, comprises the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelia as the bacterial classification that sets out; Select bright color in the present embodiment, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, and cultivate in 33 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and carries out lab scale cultivation to isolated strains, filters out two bacterial strains that mycelial growth situation is best;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.62 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties SH-2 according to mycelial growth situation.
Embodiment 3,
A disease-resistant variety selection for wild red sesame, comprises the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelia as the bacterial classification that sets out; Select bright color in the present embodiment, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, mould, Paecilomyces varioti and aspergillus spore, by this medium called after A2, and cultivate in 25 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and lab scale cultivation is carried out to isolated strains, filter out two bacterial strains that mycelial growth situation is best;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.62 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties SH-3 according to mycelial growth situation.
Embodiment 4,
A disease-resistant variety selection for wild red sesame, comprises the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelia as the bacterial classification that sets out; Select bright color in the present embodiment, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, and cultivate in 25 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and carries out lab scale cultivation to isolated strains, filters out two bacterial strains that mycelial growth situation is best;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.76 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties SH-4 according to mycelial growth situation.
Respectively the new varieties of above-described embodiment are carried out the efficiency test containing multiple miscellaneous bacterias such as Trichoderma viride, mould, aspergillus, and detect the active constituent content in 4 kinds of fruit bodys respectively, detect data as follows:
In analysis, table is known,
(1), after setting out and adding multiple miscellaneous bacteria in bacterium culture medium, the new varieties SH-3 cultivated all has higher preventive effect to Trichoderma viride and other miscellaneous bacterias, and do not add the new varieties SH-1 of other miscellaneous bacterias in medium, SH-2, SH-4 are high to Trichoderma viride preventive effect, general to other miscellaneous bacteria preventive effects, but still be better than existing kind.
(2), radiation dose is that the comprehensive resistance against diseases of SH-1, SH-2, the SH-3 in 0.62 human relations/sky and active constituent content are all higher than SH-4.
(3), Spawn incubation temperature of setting out is the SH-2 of 33 DEG C, and comparing temperature is that the anti-Trichoderma viride ability of SH-1, SH-3 and SH-4 of 25 DEG C is stronger, and active constituent content is higher.
Claims (5)
1. a disease-resistant variety selection for wild red sesame, is characterized in that, comprise the following steps:
S1, set out breeding strain: directly extract the tender somatic cell of children from the wild red sesame trophosome grown and bring out callus, then the mycelium be trained gradually, chooses pure white, healthy and strong mycelium as the bacterial classification that sets out;
S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, and cultivate in 25 DEG C of insulating boxs, the fruit body choosing robust growth carries out tissue separation, and carries out lab scale cultivation to isolated strains, filters out two bacterial strains that mycelial growth situation is best;
S3, above-mentioned two bacterial strains are trained mycelium respectively in culture bottle, and the gamma-rays accepting 0.58-0.76 human relations/sky irradiates 10 days, the high bacterial strain of the dense degree of mycelia is selected to carry out rejuvenation cultivation, separation and purification goes out different strains, and respectively different strains is trained mycelia, choose optimum bacterial strain as disease-resistant varieties according to mycelial growth situation.
2. the disease-resistant variety selection of a kind of wild red sesame as claimed in claim 1, is characterized in that: select bright color in step S1, carry out somatic cell extraction without damage by disease and insect, healthy and strong wild red sesame.
3. the disease-resistant variety selection of a kind of wild red sesame as claimed in claim 1, it is characterized in that: in step S2, will strain inoculation be set out in the PDA medium containing Trichoderma viride spore, and cultivate in the insulating box of 32-35 DEG C, the fruit body choosing robust growth carries out tissue separation, and lab scale cultivation is carried out to isolated strains, filter out two bacterial strains that mycelial growth situation is best.
4. the disease-resistant variety selection of a kind of wild red sesame as claimed in claim 1, is characterized in that: will set out strain inoculation in the PDA medium containing Trichoderma viride spore, mould, Paecilomyces varioti and aspergillus spore in step S2.
5. the disease-resistant variety selection of a kind of wild red sesame as claimed in claim 1, is characterized in that: in step S3, gamma-rays exposure intensity is 0.60-0.65 human relations/sky.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106234033A (en) * | 2016-08-26 | 2016-12-21 | 内蒙古漠菇生物科技有限公司 | Husky pestle mushroom artificial acclimation method |
| CN111887099A (en) * | 2019-11-27 | 2020-11-06 | 怀化学院 | Trichoderma viride and method for rapidly cultivating poria cocos sporocarp by trichoderma induction |
| CN113455288A (en) * | 2021-07-30 | 2021-10-01 | 龙泉市兴龙科技开发研究所 | Breeding method of disease-resistant ganoderma lucidum spore powder strain |
| CN114736808A (en) * | 2022-02-24 | 2022-07-12 | 吉林师范大学 | Method for rejuvenating trichoderma viride degenerate strain by using trace element compound amino acid |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106234033A (en) * | 2016-08-26 | 2016-12-21 | 内蒙古漠菇生物科技有限公司 | Husky pestle mushroom artificial acclimation method |
| CN111887099A (en) * | 2019-11-27 | 2020-11-06 | 怀化学院 | Trichoderma viride and method for rapidly cultivating poria cocos sporocarp by trichoderma induction |
| CN111887099B (en) * | 2019-11-27 | 2022-03-04 | 怀化学院 | Trichoderma viride and method for rapidly cultivating poria cocos sporocarp by trichoderma induction |
| CN113455288A (en) * | 2021-07-30 | 2021-10-01 | 龙泉市兴龙科技开发研究所 | Breeding method of disease-resistant ganoderma lucidum spore powder strain |
| CN114736808A (en) * | 2022-02-24 | 2022-07-12 | 吉林师范大学 | Method for rejuvenating trichoderma viride degenerate strain by using trace element compound amino acid |
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Application publication date: 20160420 |