Method for growing cyathea spinulosa sporophyte seedling cluster roots in yin

Abstract

The embodiment of the invention provides a cyathea spinulosa sporophyte cluster root preparation method, which comprises the following steps: cleaning and disinfecting feather pieces of cyathea spinulosa sporangium, taking out cyathea spinulosa spores, inoculating the cyathea spinulosa spores on a basic culture medium, culturing to obtain prothalli, transferring the prothalli to a proliferation culture medium, and carrying out proliferation induction to obtain cyathea spinulosa seedling clusters. The method comprises the following steps: adding a mixed matrix to culture a cyathea spinulosa seedling cluster, recording the number of roots, the longest root length of the roots and plant growth height index information, obtaining mixed matrix component information with the best rooting effect, and carrying out a rooting hormone test on cyathea spinulosa sporophyte seedlings under the mixed matrix component information with the best rooting effect. Compared with the prior art, the cyathea spinulosa sporophore rooting method has the advantages that cyathea spinulosa sporophore rooting workload is reduced, and the later acclimatization and transplanting survival rate is increased.

Description

Technical Field

[0001] The present application relates to the field of plant breeding, and in particular to a method for growing clustered roots of sporophyte seedlings of shade-living Alsophila spinulosa. Background Art

[0002] The spores of the shade-living tree Alsophila have a short life span, are difficult to germinate, have a long life cycle, and are difficult to resist and tolerate all adverse environmental factors, so their distribution is greatly restricted. Under modern climate conditions, the shade-living tree Alsophila can only survive in warm and humid tropical and subtropical woodlands. Once the forests where the shade-living tree Alsophila lives are destroyed, the shade-living tree Alsophila will die. Therefore, it is necessary to study the method of breeding the shade-living tree Alsophila to ensure its survival.

[0003] In the prior art, the spores of Alsophila spinulosa are disinfected and pretreated with gibberellin, collected in centrifuge tubes and stored at low temperature after drying, germinated by 75% alcohol and 4%-5% NaClO solution, and induced to root by GGB differentiation.

[0004] However, the prior art requires that the seedlings be divided into individual plants in the process of rooting induction, and the culture medium of the rooting part of the shade-growing Alsophila spinulosa must be cleaned after rooting, and finally the experimental steps such as hardening and transplanting must be carried out. Since the leaves of the shade-growing Alsophila spinulosa seedlings are papery, the seedlings are easily dehydrated and wilted during the tedious experimental process, leading to death. In the process of inducing sporophytic seedlings from the prothallus of the shade-growing Alsophila spinulosa, there is a phenomenon of symbiosis between the prothallus and the sporophytic seedlings. If the seedlings are divided into individual plants, not only the workload is large, but also the survival rate of the later hardening and transplanting will be reduced. Summary of the invention

[0005] In view of the above problems, the embodiments of the present application provide a method for growing cluster roots of sporophyte seedlings of shade-living Alsophila spinulosa, so as to solve the problems of heavy workload and low survival rate of seedling hardening and transplanting in the process of breeding shade-living Alsophila spinulosa.

[0006] In a first aspect, the present application provides a method for growing roots of Alsophila spinulosa sporophyte seedlings, comprising:

[0007] Step 1: Place the pinnae of the mature Alsophila spinulosa sporangium on filter paper to absorb the moisture on the pinnae surface, cut the pinnae, rinse with running water, place the rinsed pinnae on a workbench with alcohol, shake to remove the bubbles between the pinnae and the alcohol solution, rinse with sterile water, add HCL solution to disinfect, wash the pinnae with sterile water after disinfection, place it on sterile filter paper to absorb the moisture, then use a scalpel to open the sporangium to take out the spores, use an inoculation loop soaked in sterile water to adsorb the spores, and inoculate on a basic culture medium to culture to obtain the prothallus;

[0008] Step 2: Transferring the prothallus from the basic culture medium to the proliferation culture medium, performing prothallus proliferation and prothallus induction sporophyte seedling culture, and obtaining the shade-growing Alsophila spinulosa seedlings;

[0009] Step 3: Add the seedlings of the shade-loving Alsophila spinulosa into a culture bottle containing the mixed medium, and record the number of roots, the longest root length, and the plant growth height index information of the shade-loving Alsophila spinulosa;

[0010] Step 4: obtaining the mixed matrix component information corresponding to the shade-loving Alsophila spinulosa seedlings with the best rooting effect according to the index information, and conducting a rooting hormone test on the shade-loving Alsophila spinulosa sporophyte seedlings according to the mixed matrix component information to obtain seedling status index data.

[0011] In a possible implementation, the length of the pinna sheared in step 1 is 4-5 cm, the running water rinsing time is 30-40 min, the alcohol concentration of the workbench is 70%-75%, the number of sterile water rinsing times is 2-3 times, the concentration of the HCL solution is 0.1%, the disinfection time is 10-12 min, and the pinna is washed 3-4 times with sterile water.

[0012] In a possible embodiment, the culture conditions of the basic culture medium in step 1 are: light intensity of 2500-3000LX, light duration of 12h / d, temperature of 25±1°C, and culture time of 20 days.

[0013] In a possible embodiment, the culture conditions of the proliferation medium in step 2 are: 1 / 2 MS+KT0.3 mg / L+IBA 0.4 mg / L+sucrose 3%+agar 0.7%, and the culture time of the proliferation medium is 30 days.

[0014] In a possible embodiment, the amount of the mixed matrix in step 3 is: peat soil, peat soil and weathered soil in a ratio of 1 to 2, peat soil and weathered soil in a ratio of 1 to 1, peat soil and weathered soil in a ratio of 2 to 1, and weathered soil; after adding 30 ml of pure water to the mixed matrix, knead it into a ball by hand and fill it in a culture bottle with a filling height of 2.5 cm.

[0015] In a possible implementation, the culture bottle selected in step 3 is a plastic culture bottle with a bottom diameter of 8 cm, a height of 12 cm, and a diameter of 6.5 cm. Five groups of culture bottles are set according to the dosage of the substrate, with 10 culture bottles in each group. Two clumps of shade-growing Alsophila spinulosa seedlings are inoculated in each bottle and cultured for 30 days.

[0016] In a possible implementation manner, the mixed matrix component information corresponding to the shade-loving Alsophila spinulosa seedlings with the best rooting effect in step 4 is that the ratio of peat soil to weathered soil is 1 to 2.

[0017] In a possible embodiment, the rooting hormone test on the sporophyte seedlings of the shade-living Alsophila spinulosa is as follows: adding auxins IBA, NAA and , 16 groups of formula designs were generated through the three-factor four-level experimental method, and rooting hormone tests were carried out on the sporophyte seedlings of shade-loving Alsophila spinulosa. 16 groups of rooting information were recorded, and the 16 groups of rooting information were calculated through principal component analysis to obtain the seedling status index data.

[0018] In a possible embodiment, the concentration of the auxin IBA is 0 mg / L, 0.3 mg / L, 0.5 mg / L and 0.7 mg / L; the concentration of NAA is 0 mg / L, 0.5 mg / L, 1.0 mg / L and 1.5 mg / L; The concentrations are 0mg / L, 0.4mg / L, 0.8mg / L and 1.2mg / L.

[0019] In a possible implementation manner, the rooting information includes: the number of primary roots, the number of secondary roots, the longest root length, the base diameter, the plant height and the growth status of the plant.

[0020] The embodiment of the present application provides a method for rooting of sporophyte seedlings of shade-living Alsophila spinulosa, which comprises the following steps: cleaning and disinfecting the pinnae of the sporangium of the shade-living Alsophila spinulosa, taking out the spores of the shade-living Alsophila and inoculating them on a basic culture medium for culture to obtain prothallus, transferring the prothallus to a proliferation culture medium for proliferation induction to obtain a shade-living Alsophila seedling cluster, adding a mixed matrix to culture the shade-living Alsophila seedling cluster, recording the number of roots, the longest root length and the plant growth height index information, and obtaining the mixed matrix component information with the best rooting effect, conducting a rooting hormone test on the sporophyte seedlings of the shade-living Alsophila spinulosa under the mixed matrix component information with the best rooting effect, obtaining seedling status index data, and configuring the mixed matrix and growth hormone for rooting of the sporophyte seedlings of the shade-living Alsophila spinulosa according to the seedling status index data. Compared with the prior art, the workload of rooting of the sporophyte of the shade-living Alsophila spinulosa is reduced, and the survival rate of seedling hardening and transplanting in the later stage is improved.

[0021] In addition to the technical problems solved by the embodiments of the present application described above, the technical features that constitute the technical scheme, and the beneficial effects brought about by the technical features of the technical scheme, other technical problems that can be solved by a method for clustered roots of shade-loving Alsophila spinulosa sporophyte seedlings provided in the embodiments of the present application, other technical features included in the technical scheme, and the beneficial effects brought about by these technical features will be further described in detail in the specific implementation methods. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The accompanying drawings, which are incorporated in and constitute a part of this specification, illustrate embodiments consistent with the present application and, together with the description, serve to explain the principles of the present application.

[0023] Figure 1 A schematic diagram of the induction of sporophyte seedlings from the prothallus of the shade-loving Alsophila spinulosa provided in the examples of the present application;

[0024] Figure 2 A schematic diagram of the shade-loving Alsophila spinulosa mixed matrix provided in the embodiment of the present application for completing rooting, seedling hardening and transplanting at one time;

[0025] Figure 3 A schematic diagram of the rooting of the shade-loving Alsophila spinulosa mixed substrate provided in the examples of the present application;

[0026] Figure 4 This is a schematic diagram of the growth status of the yellowed and dead seedlings of the shade-growing Alsophila spinulosa provided in the examples of the present application.

[0027] The above drawings have shown clear embodiments of the present application, which will be described in more detail later. These drawings and text descriptions are not intended to limit the scope of the present application in any way, but to illustrate the concept of the present application to those skilled in the art by referring to specific embodiments. DETAILED DESCRIPTION

[0028] Exemplary embodiments will be described in detail herein, examples of which are shown in the accompanying drawings. When the following description refers to the drawings, the same numbers in different drawings represent the same or similar elements unless otherwise indicated. The implementations described in the following exemplary embodiments do not represent all implementations consistent with the present application. Instead, they are merely examples of devices and methods consistent with some aspects of the present application as detailed in the appended claims.

[0029] The present application embodiment provides a method for growing clustered roots of Alsophila spinulosa sporophyte seedlings, comprising:

[0030] Step 1: Place the pinnae of the mature Alsophila spinulosa sporangium on filter paper to absorb the moisture on the pinnae surface, cut the pinnae, rinse with running water, place the rinsed pinnae on a workbench with alcohol, shake to remove the bubbles between the pinnae and the alcohol solution, rinse with sterile water, add HCL solution to disinfect, wash the pinnae with sterile water after disinfection, place it on sterile filter paper to absorb the moisture, then use a scalpel to open the sporangium to take out the spores, use an inoculation loop soaked in sterile water to adsorb the spores, and inoculate on a basic culture medium to culture to obtain the prothallus;

[0031] Step 2: Transferring the prothallus from the basic culture medium to the proliferation culture medium, performing prothallus proliferation and prothallus induction sporophyte seedling culture, and obtaining the shade-growing Alsophila spinulosa seedlings;

[0032] Step 3: Add the seedlings of the shade-loving Alsophila spinulosa into a culture bottle containing the mixed medium, and record the number of roots, the longest root length, and the plant growth height index information of the shade-loving Alsophila spinulosa;

[0033] Step 4: According to the index information, the mixed matrix component information corresponding to the shade-loving Alsophila spinulosa seedlings with the best rooting effect is obtained, and the rooting hormone test is performed on the shade-loving Alsophila spinulosa sporophyte seedlings according to the mixed matrix component information to obtain the seedling status index data.

[0034] Among them, in step 1, the pinnae of mature Alsophila spinulosa sporangium are first placed on filter paper to absorb the moisture on the surface of the pinnae, and then the pinnae are cut into 4-5 cm in length, and the cutting length is preferably such that the sporangium is not cut, and the pinnae are rinsed with running water for 30-40 minutes. The rinsed pinnae are placed in an ultra-clean workbench with 70-75% alcohol, and the bubbles between the pinnae and the alcohol solution are shaken to expel, and the pinnae are rinsed with sterile water for 2-3 times, and then 0.1% HCL solution is added for disinfection for 10-12 minutes, and then washed with sterile water for 3-4 times after disinfection, and the disinfected pinnae with the spores of Alsophila spinulosa are placed on sterile filter paper to absorb the moisture on both sides of the pinnae, and the sporangium is peeled open with a scalpel to remove the spores, and the spores are adsorbed with an inoculation loop soaked in sterile water, and inoculated on a DCR basic culture medium without adding any exogenous hormones.

[0035] In this embodiment, the culture conditions of the basic culture medium in step 1 are: light intensity of 2500-3000LX, light duration of 12h / d, temperature of 25±1°C, and culture time of 20 days.

[0036] Among them, the culture conditions of the proliferation medium in step 2 are: 1 / 2 MS+KT 0.3 mg / L+IBA 0.4 mg / L+sucrose 3%+agar 0.7%, and the culture time of the proliferation medium is 30 days.

[0037] Figure 1 This is a schematic diagram of the induction of sporophyte seedlings from the prothallus of the shade-loving Alsophila spinulosa provided in the examples of the present application.

[0038] In one embodiment of the present application, the amount of the mixed matrix in step 3 is divided into 5 groups of ratios:

[0039] 1. Peat soil.

[0040] 2. The ratio of peat soil to weathered soil is 1 to 2.

[0041] 3. The ratio of peat soil to weathered soil is 1:1.

[0042] 4. The ratio of peat soil to weathered soil is 2 to 1.

[0043] 5. Weathered soil.

[0044] After adding 30 ml of pure water to the mixed matrix, knead it into a ball by hand, fill it to a height of 2.5 cm in a culture bottle.

[0045] Among them, the culture bottle is a plastic culture bottle with a bottom diameter of 8 cm, a height of 12 cm, and a diameter of 6.5 cm. According to the 5 groups of mixed substrate ratios, 10 culture bottles are set for each group of ratios, and 2 clumps of Alsophila spinulosa seedlings are inoculated in each bottle for 30 days. After 30 days, the number of roots, the longest root length, and the growth height of the plants of Alsophila spinulosa were counted. The experimental results are shown in Table 1.

[0046] Table 1 Rooting statistics of the mixed substrate of the sporophyte seedlings of Alsophila spinulosa

[0047]

[0048] Among them, different lowercase letters represent significant differences, P < 0.05. According to the experimental results in Table 1, the mixed matrix peat soil: weathered soil = 1:2 combination had the best effect. The number of clustered roots of the shade-living Alsophila spinulosa sporophyte seedlings reached 3.15, the longest root length was 17.1 mm, and the plant growth height was 28.5 mm.

[0049] Figure 2 This is a schematic diagram of the shade-loving Alsophila spinulosa mixed matrix provided in the embodiment of the present application for completing rooting, seedling hardening and transplanting at one time.

[0050] In one embodiment of the present application, the rooting hormone test on the sporophyte seedlings of the shade-living Alsophila spinulosa in step 4 is as follows: adding auxins IBA, NAA and , 16 groups of formula designs were generated through the three-factor four-level experimental method, and rooting hormone tests were carried out on the sporophyte seedlings of shade-loving Alsophila spinulosa. 16 groups of rooting information were recorded, and the 16 groups of rooting information were calculated through principal component analysis to obtain the seedling status index data.

[0051] In this embodiment, the three factors in the three-factor four-level test method include: auxin IBA, NAA and The selection of culture bottles, the capacity of the mixed matrix, and the amount of hormone solution are the same as the capacity of pure water added and the state of the mixed matrix in step 3.

[0052] In this embodiment, the concentration of the auxin IBA is 0 mg / L, 0.3 mg / L, 0.5 mg / L and 0.7 mg / L; the concentration of NAA is 0 mg / L, 0.5 mg / L, 1.0 mg / L and 1.5 mg / L; The concentrations are 0mg / L, 0.4mg / L, 0.8mg / L and 1.2mg / L.

[0053] Among them, auxin IBA, NAA and The rooting hormone factor levels are shown in Table 2.

[0054] Table 2 Rooting hormone factor levels of Alsophila spinulosa sporophyte seedlings

[0055]

[0056] Among them, according to the three-factor four-level design, the mixed matrix was peat soil: weathered soil = 1:2, and different hormone types and concentrations were added. The 16 groups of treatment formulas were designed as follows:

[0057] 1. Peat soil: weathered soil = 1:2

[0058] 2. Peat soil: weathered soil = 1:2 + NAA 0.5mg / L+ 0.4mg / L

[0059] 3. Peat soil: weathered soil = 1:2 + NAA 1.0mg / L+ 0.8mg / L

[0060] 4. Peat soil: weathered soil = 1:2 + NAA 1.5mg / L+ 1.2mg / L

[0061] 5. Peat soil: weathered soil = 1:2 + IBA 0.3mg / L+ 0.4mg / L

[0062] 6. Peat soil: weathered soil = 1:2 + IBA 0.3mg / L + NAA 0.5mg / L

[0063] 7. Peat soil: weathered soil = 1:2 + IBA 0.3mg / L + NAA 1.0mg / L + 1.2mg / L

[0064] 8. Peat soil: weathered soil = 1:2 + IBA 0.3mg / L + NAA 1.5mg / L + 0.8mg / L

[0065] 9. Peat soil: weathered soil = 1:2 + IBA 0.5mg / L+ 0.8mg / L

[0066] 10. Peat soil: weathered soil = 1:2 + IBA 0.5mg / L + NAA 0.5mg / L + 1.2mg / L

[0067] 11. Peat soil: weathered soil = 1:2 + IBA 0.5mg / L + NAA 1.0mg / L

[0068] 12. Peat soil: weathered soil = 1:2 + IBA 0.5mg / L + NAA 1.5mg / L + 0.4mg / L

[0069] 13. Peat soil: weathered soil = 1:2 + IBA 0.7mg / L+ 1.2mg / L

[0070] 14. Peat soil: weathered soil = 1:2 + IBA 0.7mg / L + NAA 0.5mg / L + 0.8mg / L

[0071] 15. Peat soil: weathered soil = 1:2 + IBA 0.7mg / L + NAA 1.0mg / L + 0.4mg / L

[0072] 16. Peat soil: weathered soil = 1:2 + IBA 0.7mg / L + NAA 1.5mg / L

[0073] In this embodiment, in the 16 groups of formula design, 10 culture bottles are set in each group, and 2 seedlings are inoculated in each bottle. After 30 days of cultivation, the rooting information of the 16 groups is counted, and the rooting information includes: the number of primary roots, the number of secondary roots, the longest root length, the base diameter, the plant height and the plant growth status. The rooting statistics of the 16 groups of shade-growing Alsophila spinulosa sporophyte seedling rooting hormone combinations are shown in Table 3.

[0074] Table 3 Rooting statistics of the rooting hormone combination of Alsophila spinulosa seedlings

[0075]

[0076] Among them, health rate = 1-number of yellowed and dead plants / number of treated plants*100%; different lowercase letters represent significant differences, P < 0.05.

[0077] As shown in Table 3, under the condition of different hormone combinations in the same mixed matrix, the best performance in the number of primary roots was in treatment 3, with an average number of 7.1 roots, which was higher than the other 15 treatments. The best performance in the number of secondary roots was in treatment 3, with an average number of 1.3 roots, which was higher than the other treatments except treatments 11 and 12. The best performance in the longest root length was in treatment 1, with an average length of 29.35 mm, significantly higher than treatments 9 and 15. The best performance in the base diameter was in treatment 3, with an average length of 6.08 mm, which was higher than the other treatments except treatment 6. The best performance in plant height was in treatment 2, with an average length of 41.51 mm, which was significantly higher than the other treatments except treatments 3, 5, 6, 15 and 16. The best performance in health rate was in treatment 3, with an average health rate of 90%, which was significantly higher than the other treatments except treatments 2, 12 and 16.

[0078] Figure 3 This is a schematic diagram of the rooting of the shade-loving Alsophila spinulosa mixed matrix provided in the examples of the present application.

[0079] Figure 4 This is a schematic diagram of the growth status of the yellowed and dead seedlings of the shade-growing Alsophila spinulosa provided in the examples of the present application.

[0080] In this embodiment, principal component analysis was performed on the six types of rooting information indicators in Table 3, and the comprehensive scores of the main indicators were calculated. The scores of the 16 treatment groups are shown in Table 4.

[0081] Table 4 Score table of rooting hormone combinations for seedling status of shade-loving Alsophila spinulosa sporophyte seedlings

[0082]

[0083] Referring to Table 3 and Table 4, the rooting medium for the sporophyte seedlings of the shade-growing Alsophila spinulosa should be selected with peat soil: weathered soil = 1:2 for the best rooting effect. The number of clustered roots of the sporophyte seedlings of the shade-growing Alsophila spinulosa reached 3.15, the longest root length was 17.1mm, and the plant growth height was 28.5mm; in the experiment of different hormone combinations in the same mixed matrix, the number of rooting roots of the first-level roots, the number of rooting roots of the second-level roots, the base diameter and the plant growth status of the combination No. 3 treatment were better than the other treatment combination levels. Its various indicators are: the number of first-level roots reached 7.1, the number of second-level roots was 1.3, the base diameter was 6.08mm, and the proportion of green plants in the plant growth status was 90%. Through the second rooting test of the shade-growing Alsophila spinulosa, it was found that the number of first-level roots in the second rooting test was 3.95 higher than that in the first rooting test; the longest root length was 10.56mm higher; and the plant growth height was 11.6mm higher. In the mixed matrix peat soil: weathered soil = 1:2, add NAA1.0mg / L+ The interaction between auxins under the 0.8 mg / L treatment can better promote the rooting and plant growth of shade-loving Alsophila spinulosa, and the rooting indicators can achieve significant effects under the appropriate mixed substrate ratio.

[0084] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit it. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that they can still modify the technical solutions described in the aforementioned embodiments, or replace some or all of the technical features therein with equivalents. However, these modifications or replacements do not cause the essence of the corresponding technical solutions to deviate from the scope of the technical solutions of the embodiments of the present invention.

Claims

1. A method for growing roots of Alsophila spinulosa sporophyte seedlings, characterized in that: include: Step 1: Place the pinnae of the mature Alsophila spinulosa sporangium on filter paper to absorb the moisture on the pinnae surface, cut the pinnae, rinse with running water, place the rinsed pinnae on a workbench with alcohol, shake to remove the bubbles between the pinnae and the alcohol solution, rinse with sterile water, add HCL solution to disinfect, wash the pinnae with sterile water after disinfection, place it on sterile filter paper to absorb the moisture, then use a scalpel to open the sporangium to take out the spores, use an inoculation loop soaked in sterile water to adsorb the spores, and inoculate on a basic culture medium to culture to obtain the prothallus; Step 2: Transferring the prothallus from the basic culture medium to the proliferation culture medium, performing prothallus proliferation and prothallus induction sporophyte seedling culture, and obtaining the shade-growing Alsophila spinulosa seedlings; Step 3: Add the seedlings of the shade-loving Alsophila spinulosa into a culture bottle containing the mixed medium, and record the number of roots, the longest root length, and the plant growth height index information of the shade-loving Alsophila spinulosa; Step 4: obtaining the mixed matrix component information corresponding to the shade-loving Alsophila spinulosa seedlings with the best rooting effect according to the index information, and conducting a rooting hormone test on the shade-loving Alsophila spinulosa sporophyte seedlings according to the mixed matrix component information to obtain seedling status index data.

2. The method according to claim 1, characterized in that The pinnae cut in step 1 are 4-5 cm in length, the running water rinsing time is 30-40 min, the alcohol concentration of the workbench is 70%-75%, the sterile water rinsing times are 2-3 times, the concentration of the HCL solution is 0.1%, the disinfection time is 10-12 min, and the pinnae are washed 3-4 times with sterile water.

3. The method according to claim 1, characterized in that The culture conditions of the basic culture medium in step 1 are: light intensity of 2500-3000LX, light duration of 12h / d, temperature of 25±1°C, and culture time of 20 days.

4. The method according to claim 1, characterized in that: The culture conditions of the proliferation medium in step 2 are: 1 / 2 MS+KT 0.3 mg / L+IBA 0.4 mg / L+sucrose 3%+agar 0.7%, and the culture time of the proliferation medium is 30 days.

5. The method according to claim 1, characterized in that The dosage of the mixed matrix in the step 3 is: peat soil, peat soil and weathered soil in a ratio of 1 to 2, peat soil and weathered soil in a ratio of 1 to 1, peat soil and weathered soil in a ratio of 2 to 1, and weathered soil; the mixed matrix is ​​added with 30 ml of pure water and kneaded into a ball by hand, and the filling height is 2.5 cm and filled in a culture bottle.

6. The method according to claim 5, characterized in that The culture bottles selected in step 3 are plastic culture bottles with a bottom diameter of 8 cm, a height of 12 cm, and a diameter of 6.5 cm. Five groups of culture bottles are set according to the dosage of the substrate, with 10 culture bottles in each group. Two clumps of shade-growing Alsophila spinulosa seedlings are inoculated in each bottle and cultured for 30 days.

7. The method according to claim 1, characterized in that The mixed matrix component information corresponding to the shade-loving Alsophila spinulosa seedlings with the best rooting effect in step 4 is that the ratio of peat soil to weathered soil is 1 to 2.

8. The method according to any one of claims 1 to 7, characterized in that: The rooting hormone test on the sporophyte seedlings of the shade-living Alsophila spinulosa is as follows: the auxins IBA, NAA and , 16 groups of formula designs were generated through the three-factor four-level experimental method, and rooting hormone tests were carried out on the sporophyte seedlings of shade-loving Alsophila spinulosa. 16 groups of rooting information were recorded, and the 16 groups of rooting information were calculated through principal component analysis to obtain the seedling status index data.

9. The method according to claim 8, characterized in that The concentrations of the auxin IBA are 0 mg / L, 0.3 mg / L, 0.5 mg / L and 0.7 mg / L; the concentrations of NAA are 0 mg / L, 0.5 mg / L, 1.0 mg / L and 1.5 mg / L; The concentrations are 0mg / L, 0.4mg / L, 0.8mg / L and 1.2mg / L.

10. The method according to claim 8, characterized in that The rooting information includes: the number of primary roots, the number of secondary roots, the longest root length, the base diameter, the plant height and the growth status of the plant.

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