CN113940271A - Breeding and propagating method of cold-resistant stylosanthes guianensis - Google Patents
Breeding and propagating method of cold-resistant stylosanthes guianensis Download PDFInfo
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Abstract
The invention belongs to the technical field of plant breeding, and particularly relates to a breeding and propagation method of cold-resistant stylosanthes guianensis. Adopts Oxley stylosanthes guianensis with strong cold resistance and high-yield and high-quality stylosanthes guianensis as hybridization parents, the seeds are pretreated by low-frequency current treatment and bacterial liquid soaking treatment to stimulate the activity of the seeds, the water solution of cane sugar and boric acid is sprayed on the styluses of the styluses in the bud period to improve the vitality and the durability of pollen and help to promote the hybridization among the varieties of the styluses of different varieties, and then, the growth bacterial liquid is adopted to promote the growth and development of the seedlings, the obtained cold-resistant and high-yield stylosanthes guianensis plants have higher quality than the parent plants, the method can be used for supplying green feed for livestock and poultry in south China, processing grass meal and producing compound feed, intercropping and covering in forest orchards, improving grazing land and the like, and has important significance for developing the grazing industry by introducing the stylosanthes guianensis into the north due to cold resistance and high quality.
Description
Technical Field
The invention belongs to the technical field of plant breeding, and particularly relates to a breeding and propagation method of cold-resistant stylosanthes guianensis.
Background
Stylosanthes guianensis is a perennial legume grass native to Latin America, widely distributed in tropical and subtropical regions, about 50 species worldwide. Stylosanthes guianensis prefers high-temperature and high-humidity environment, and has become excellent pasture widely planted in tropical and subtropical regions in south China due to the characteristics of high yield, good quality, strong adaptability and the like.
Among them, stylosanthes guianensis shows extremely excellent growth characteristics in the aspects of stress resistance and adaptability, is outstanding in the aspects of waterlogging tolerance, soil barren tolerance, acid resistance, disease and insect resistance and the like, and still shows higher production performance in middle and low fertility areas. At present, stylosanthes guianensis becomes important high-quality pasture in southern areas of China, and is an important component of high-quality forage grass and feed resources for livestock and poultry cultivation.
However, in production and application, the current columella pulchrum main-pushing variety is difficult to overwinter in the central and north parts of the south China, the planting and popularization of the columella pulchrum is limited to a great extent, and the problem of safe overwinter of the columella pulchrum becomes the primary problem of production and popularization and application of the columella pulchrum in the south China.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a breeding method of cold-resistant stylosanthes guianensis, and on the basis, stylosanthes guianensis plants with strong cold resistance, high yield and high quality are obtained.
The technical content of the invention is as follows:
the invention provides a breeding method of cold-resistant stylosanthes guianensis, which comprises the following steps:
1) selecting Oxley stylosanthes guianensis as a female parent and stylosanthes guianensis as a male parent;
2) seed pretreatment: the Oxley stylosanthes guianensis seeds are subjected to low-frequency current treatment and soaked in streptomyces clausii (product number: XK-GDJZ-0440) solution, so that the activity of the seeds is activated, the germ infection is prevented, the immunological competence of the seeds is improved, and the germination of the seeds and the growth of seedlings are promoted. Selecting strong plants;
soaking the stylosanthes guianensis seeds in a mixed solution of trichoderma reesei, humus and an abscisic acid aqueous solution, alternately performing ice bath and hot water bath, combining the trichoderma reesei and the humus with phytohormone abscisic acid, promoting genetic modification of the seeds, improving the bioactivity of the seeds, and promoting germination of the seeds and growth of seedlings;
3) and (3) treating planting soil: red sandy soil, pH 5-6, organic matter content 1-1.5%, total nitrogen content 0.02-0.05%, total phosphorus content 0.05-0.08%, total potassium content 0.4-0.6%, without irrigation;
4) carrying out field planting on the pretreated Oxley stylosanthes guianensis seeds and the stylosanthes guianensis seeds, selecting strong plants, spraying a sucrose + boric acid aqueous solution on stylosanthes guianensis inflorescences in the bud period, improving the vitality and the durability of pollen and facilitating the hybridization among the stylosanthes guianensis varieties of different varieties;
performing artificial emasculation on Oxley stylosanthes guianensis in the bud stage, performing artificial pollination on pollen of other stylosanthes guianensis in the bud stage, hybridizing the parents of the Oxley stylosanthes guianensis and the other stylosanthes guianensis to obtain an F1 generation, and selecting the seedling with better growth vigor as the cold-resistant stylosanthes guianensis seedling;
5) transplanting selected cold-resistant stylosanthes guianensis seedlings, 1-2 seedlings are transplanted in each hole, and uniformly spraying growth bacterium liquid on the leaves of the seedlings for natural growth;
6) and when branches grow out from the cold-resistant stylobactrium roseum seedlings, selecting the branches as seedlings, and carrying out cultivation and planting to obtain the cold-resistant stylobactrium roseum plants.
Step 2), the concentration of the streptomyces meretrix bacterial liquid is 0.3-0.5%, and the streptomyces meretrix bacterial liquid is a mixed liquid of streptomyces meretrix bacterial powder and deionized water;
the solid-to-liquid ratio of the mixed solution of trichoderma reesei, humus and abscisic acid aqueous solution is (3-5): (15-20) g/mL;
the mass ratio of the trichoderma reesei to the humus is 1: (70-85), wherein the concentration of the abscisic acid aqueous solution is 25-35%.
And 2) treating the low-frequency current for 8-12 min by adopting 40-50 Hz for 3-4 times at intervals of 20-30 min.
The growth bacterial liquid in the step 3) is a mixed bacterial liquid of the Geospora and the Trichoderma harzianum T-22, the concentration is 0.3-0.5%, and the mass ratio of the powder of the Geospora and the Trichoderma harzianum T-22 is (1-2): (2-5).
In the aqueous solution of sucrose and boric acid in the step 4), the concentration of sucrose is 10-20%, and the concentration of boric acid is 0.01-0.02%.
Step 5) the specific operation is as follows: selecting branches with the length of 30-35 cm as seedlings, planting in 3-ten days of the month, soaking grass seedlings into roots for 12-24 hours before planting the seedlings, burying soil with the depth of 10-15 cm, vertically transplanting, pouring root fixing water, and then performing field management and disease control.
The areas of hybridization in the step 4), transplantation in the step 5) and cultivation in the step 6) are all stylosanthes of south China.
The Stylosanthes guianensis Oxley has excellent cold resistance, but the stems and the thin plants are short, the yield is low, and other Stylosanthes guianensis such as Stylosanthes gracilis, Stylosanthes guianensis 2, Stylosanthes guianensis 5 or Stylosanthes guianensis are high-yield and high-quality Stylosanthes guianensis, but the cold resistance is poor, so that the Stylosanthes guianensis Oxley cannot safely live through the winter.
The invention has the following beneficial effects:
according to the alternate propagation method of the cold-resistant stylosanthes guianensis, Oxley stylosanthes guianensis with strong cold resistance and high-yield and high-quality stylosanthes guianensis are adopted as hybrid parents, and the seeds are subjected to low-frequency current treatment and bacterial liquid soaking treatment to stimulate the activity of the seeds so as to promote the germination of the seeds and the growth of seedlings; spraying aqueous solution of sucrose and boric acid on stylosanthes guianensis inflorescences in the bud stage for spraying treatment, so that the vigor and the durability of pollen are improved, the hybridization among stylosanthes guianensis varieties of different varieties is facilitated, and the problem that the natural outcrossing rate of Oxley stylosanthes guianensis and other stylosanthes guianensis is only 1% is solved;
after the Oxley stylosanthes guianensis and the stylosanthes guianensis are hybridized, growth bacteria liquid is adopted to promote the growth and development of seedlings, 20-30% of stylosanthes guianensis strains in the first generation of seedlings are shown as the stylosanthes guianensis, and the growth speed is high; selecting seedlings to transplant, namely carrying out vegetative propagation to obtain cold-resistant thin-stem stylosanthes guianensis with stable genetic character;
compared with Oxley stylosanthes guianensis, the yield, the plant height, the leaf area and other agronomic characters of the cold-resistant stylosanthes guianensis obtained by the invention are basically consistent with those of other high-yield and high-quality stylosanthes guianensis varieties, and meanwhile, the cold-resistant stylosanthes guianensis has stronger cold resistance and obvious heterosis. The obtained cold-resistant and high-yield stylosanthes guianensis plants have higher quality than parents, contain higher content of nutrient elements, can be used for supplying green feed for livestock in south China, processing grass meal and producing compound feed, intercropping and covering in forest orchards, improving the popularization and application in the aspects of grazing land and the like, and have important significance for exploiting the breeding industry by the stylosanthes guianensis due to the cold-resistant and high-quality of the stylosanthes guianensis.
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FIG. 1 is a schematic diagram of the breeding and propagation of the cold-resistant stylosanthes guianensis.
Detailed Description
The present invention is described in further detail in the following detailed description with reference to specific embodiments, which are intended to be illustrative only and not to be limiting of the scope of the invention, as various equivalent modifications of the invention will become apparent to those skilled in the art after reading the present invention and are intended to be included within the scope of the appended claims.
All the raw materials and reagents of the invention are conventional market raw materials and reagents unless otherwise specified.
The whole process of the breeding field of the stylosanthes guianensis is the stylosanthes guianensis pasture in south China, the original Qujiang demonstration pasture in north Guangdong is selected in the embodiment of the invention, the time of parent hybridization is 11-12 months, and the planting area is set to be 12m2。
Example 1
A method for breeding cold-resistant stylosanthes guianensis comprises the following steps:
1) selecting Oxley stylosanthes guianensis seeds as a female parent and stylosanthes guianensis as a male parent;
2) seed pretreatment: carrying out 40Hz low-frequency current treatment on Oxley stylosanthes guianensis seeds for 12min for 4 times at intervals of 20min, and then soaking the Oxley stylosanthes guianensis seeds in a mixed solution of streptomyces clausii powder and deionized water, wherein the concentration of the Oxley stylosanthes guianensis seeds is 0.5%;
soaking the stylosanthes guianensis seeds in a mixed solution of trichoderma reesei, humus and an abscisic acid aqueous solution, wherein the solid-to-liquid ratio is 3:15g/mL, and ice bath and hot water bath are alternately carried out for 10 minutes respectively and 3 times;
wherein the mass ratio of the trichoderma reesei to the humus is 1: 75, the concentration of the abscisic acid aqueous solution is 30 percent;
3) and (3) treating planting soil: the pH value is 6, the organic matter content is 1.5%, the total nitrogen content is 0.5%, the total potassium content is 0.6%, and irrigation is not performed;
4) carrying out field planting on the pretreated Oxley stylosanthes guianensis seeds and the stylosanthes guianensis seeds, selecting strong plants, and spraying aqueous solution of 15 percent of sucrose and 0.01 percent of boric acid on stylosanthes guianensis inflorescences in the bud period;
performing artificial emasculation on Oxley stylosanthes guianensis in the bud stage, performing artificial pollination on the pollen of the stylosanthes guianensis in the bud stage, performing hybridization to obtain F1 generations, and selecting the seedling with better growth vigor as the cold-resistant stylosanthes guianensis seedling;
5) transplanting selected cold-resistant stylosanthes guianensis seedlings, 1-2 seedlings are transplanted in each hole, and uniformly spraying growth bacterium liquid on the leaves of the seedlings for natural growth;
the growth bacterial liquid is a mixed bacterial liquid of the Geospora and the Trichoderma harzianum T-22, the concentration is 0.5%, and the mass ratio of the powder of the Geospora and the Trichoderma harzianum T-22 is 2: 5;
6) when branches grow out from the cold-resistant stylobate flower and grass seedlings, selecting the branches as seedlings, and planting: selecting branches with the length of 30cm as seedlings, planting in 3 middle ten days, soaking grass seedlings into roots for 18 hours before planting the seedlings, burying soil with the depth of 12cm, vertically transplanting, watering root fixing water, and then performing field management and disease control to obtain cold-resistant stylobate flower and grass plants, wherein the process is shown in figure 1 and is a schematic diagram of the breeding and propagation process of the cold-resistant stylobate flower and grass.
Field management: after planting, determining whether watering is needed according to weather conditions, removing impurities once after two weeks, enabling plants to grow, if dead seedlings are found, timely supplementing the seedlings, and then carrying out daily management such as intertillage, weeding, fertilization and the like;
and (3) pest control: rarely finds plant diseases and insect pests, but in the grassland planted for many years, if anthrax infection is found, the medicine is required to be controlled;
example 2
A method for breeding cold-resistant stylosanthes guianensis comprises the following steps:
1) selecting Oxley stylosanthes guianensis seeds as a female parent and stylosanthes guianensis as a male parent;
2) seed pretreatment: carrying out low-frequency current treatment on Oxley stylosanthes guianensis seeds at 45Hz for 10min for 3 times at intervals of 25min, and then soaking the seeds in a mixed solution of streptomyces clausii powder and deionized water, wherein the concentration of the seeds is 0.4%;
soaking the stylosanthes guianensis seeds in a mixed solution of trichoderma reesei, humus and an abscisic acid aqueous solution, wherein the solid-to-liquid ratio is 4:17g/mL, and ice bath and hot water bath are alternately carried out for 8 minutes respectively and 3 times;
wherein the mass ratio of the trichoderma reesei to the humus is 1: 70, the concentration of the abscisic acid aqueous solution is 25 percent;
3) and (3) treating planting soil: the pH value is 5, the organic matter content is 1.3%, the total nitrogen content is 0.2%, the total potassium content is 0.5%, and irrigation is not performed;
4) carrying out field planting on the pretreated Oxley stylosanthes guianensis seeds and the stylosanthes guianensis seeds, selecting strong plants, and spraying aqueous solution of 10% sucrose and 0.01% boric acid to stylosanthes guianensis inflorescences in the bud period;
performing artificial emasculation on Oxley stylosanthes guianensis in the bud stage, performing artificial pollination on the pollen of the stylosanthes guianensis in the bud stage, performing hybridization to obtain F1 generations, and selecting the seedling with better growth vigor as the cold-resistant stylosanthes guianensis seedling;
5) transplanting selected cold-resistant stylosanthes guianensis seedlings, 1-2 seedlings are transplanted in each hole, and uniformly spraying growth bacterium liquid on the leaves of the seedlings for natural growth;
the growth bacterial liquid is a mixed bacterial liquid of the Geospora and the Trichoderma harzianum T-22, the concentration is 0.4%, and the mass ratio of the powder of the Geospora and the Trichoderma harzianum T-22 is 1: 3;
6) when branches grow out from the cold-resistant stylobate flower and grass seedlings, selecting the branches as seedlings, and planting: selecting branches with the length of 33cm as seedlings, planting in 3 middle ten days, soaking grass seedlings into roots for 12 hours before planting the seedlings, burying soil with the depth of 10cm, vertically transplanting, watering root fixing water, and then performing field management and disease control to obtain the cold-resistant stylobate flower and grass plants.
Example 3
A method for breeding cold-resistant stylosanthes guianensis comprises the following steps:
1) selecting Oxley stylosanthes guianensis seeds as a female parent and stylosanthes guianensis as a male parent;
2) seed pretreatment: carrying out 50Hz low-frequency current treatment on Oxley stylosanthes guianensis seeds for 12min for 3 times at intervals of 20min, and then soaking the Oxley stylosanthes guianensis seeds in a mixed solution of streptomyces clausii powder and deionized water, wherein the concentration of the Oxley stylosanthes guianensis seeds is 0.3%;
soaking the stylosanthes guianensis seeds in a mixed solution of trichoderma reesei, humus and an abscisic acid aqueous solution, wherein the solid-to-liquid ratio is 5:20g/mL, and ice bath and hot water bath are alternately carried out for 8 minutes respectively and 3 times;
wherein the mass ratio of the trichoderma reesei to the humus is 1:85, and the concentration of the abscisic acid aqueous solution is 35%;
3) and (3) treating planting soil: the pH value is 5, the organic matter content is 1%, the total nitrogen content is 0.4%, the total potassium content is 0.4%, and irrigation is not needed;
4) carrying out field planting on the pretreated Oxley stylosanthes guianensis seeds and the stylosanthes guianensis seeds, selecting strong plants, and spraying an aqueous solution of 20 percent of sucrose and 0.02 percent of boric acid on stylosanthes guianensis inflorescences in the bud period;
performing artificial emasculation on Oxley stylosanthes guianensis in the bud stage, performing artificial pollination on the pollen of the stylosanthes guianensis in the bud stage, performing hybridization to obtain F1 generations, and selecting the seedling with better growth vigor as the cold-resistant stylosanthes guianensis seedling;
5) transplanting selected cold-resistant stylosanthes guianensis seedlings, 1-2 seedlings are transplanted in each hole, and uniformly spraying growth bacterium liquid on the leaves of the seedlings for natural growth;
the growth bacterial liquid is a mixed bacterial liquid of the Geospora and the Trichoderma harzianum T-22, the concentration is 0.3 percent, and the mass ratio of the powder of the Geospora and the Trichoderma harzianum T-22 is 1: 2;
6) when branches grow out from the cold-resistant stylobate flower and grass seedlings, selecting the branches as seedlings, and planting: selecting branches with the length of 35cm as seedlings, planting in 3 middle ten days, soaking grass seedlings into roots for 24 hours before planting the seedlings, burying soil with the depth of 15cm, vertically transplanting, watering root fixing water, and then performing field management and disease control to obtain the cold-resistant stylobate flower and grass plants.
Comparative example 1
The control group of example 1, the pretreatment of the parental seeds was eliminated (i.e., step 2).
Comparative example 2
The control group of example 1, the spraying of the aqueous solution of sucrose + boric acid (step 4) on the young seedlings at the bud stage was cancelled.
Comparative example 3
The control group of example 1, the growth and development treatment of the seedlings (i.e., the use of growth medium in step 5) was omitted.
Comparative example 4
The control group of example 1, the transplantation of shoots was cancelled (i.e., step 6).
The following is a variety comparison test between the cold-resistant Stylosanthes guianensis plants obtained by the present invention and the existing Stylosanthes guianensis plants.
The altitude of the selected test pasture is 40m, the annual average temperature is 22 ℃, the annual average rainfall is 1600-2000 mm, the sunshine hours are 1945h, and the soil is red sandy loam.
1. Growth rate
The cold-resistant stylosanthes guianensis plant planted in the example 1, the stylosanthes guianensis of the comparative example 2, the stylosanthes guianensis of the comparative example 3, the stylosanthes guianensis of the comparative example 4, the Oxley stylosanthes guianensis, the stylosanthes guianensis of the gram, the stylosanthes guianensis of the heat grinding No. 2, the stylosanthes guianensis of the heat grinding No. 5 and the branches with the hooked stylosanthes guianensis (Roche) are vertically inserted into soil, and the growth height and the growth speed are recorded, and the results are shown in the table 1.
TABLE 1 height comparison (cm) of stylosanthes guianensis plants
The cold-resistant stylosanthes guianensis obtained by the method starts to grow 15 days after transplantation, and the later growth speed is higher. Compared with the parent, the growth rate of the cold-resistant stylosanthes guianensis is obviously improved, and the growth performance is stronger. Compared with the stylosanthes guianensis plant of the comparative example 1, except that the seed hybridization success rate of the comparative example 1 is lower, the growth speed and the height of the plant are almost the same;
compared with the stylosanthes guianensis plant in the comparative example 2 and the stylosanthes guianensis plant in the comparative example 3, the stylosanthes guianensis plant in the example 1 has obvious growth advantages, and the adopted growth bacterial liquid and the transplanting measures have obvious effects on the growth of the stylosanthes guianensis.
2. Nutritional levels
After the stylosanthes guianensis grows for 150 days, selecting stylosanthes guianensis plants of each group, detecting the nutrient content and the content in branches of the stylosanthes guianensis plants after drying treatment, and averaging the values to obtain the results shown in the following table.
TABLE 2 nutrient composition of Stylosanthes guianensis plants (oven dried,%)
The table shows that the cold-resistant stylosanthes guianensis obtained by the method is excellent in nutritional quality, crude protein and crude fat of the stylosanthes guianensis are improved by 15-50%, crude fiber of the stylosanthes guianensis is improved by 5-20%, nitrogen-free extract of the stylosanthes guianensis is improved by 5-10%, calcium of the stylosanthes guianensis is improved by 50-70%, phosphorus of the stylosanthes guianensis is improved by 70-150%, and the levels of nutritional ingredients of the stylosanthes guianensis, parent stylosanthes guianensis and other common stylosanthes guianensis of the comparative example are all higher than those of the stylosanthes guianensis, parent stylosanthes guianensis and other common stylosanthes guianensis of the comparative example.
3. Grass yield
TABLE 3 first mowing yield (kg) of stylosanthes guianensis
TABLE 4 grass yield (kg) for the second mowing of stylosanthes guianensis
As can be seen from the table, the grass yield of the cold-resistant stylosanthes guianensis obtained by the method is highest no matter the stylosanthes guianensis is mown for the first time or the second time, and the grass yield is high, so that the cold-resistant stylosanthes guianensis has excellent growth characteristics.
The grass yield of comparative example 2 was inferior to that of example 1, and it was found that the pretreatment of seeds, the growth treatment of seedlings and the transplanting culture of plants by the present invention had an accelerating effect on the growth of Stylosanthes guianensis.
4. Overwintering rate
TABLE 5 questionnaire of overwintering rate of Styrax ramosissimum (2020.12 ~ 2021.03)
As can be seen from the table, the research on the survival plants of the stylosanthes guianensis from 12 months in 2020 to 03 months in 2021 shows that the invention obtains the Oxley stylosanthes guianensis with excellent cold resistance, and the overwintering rate reaches 96 percent and is basically consistent with that of the female parent.
The overwintering rate of the stylosanthes guianensis obtained in the comparative examples 1-3 is slightly reduced, and the pretreatment of seeds, the growth treatment of seedlings and the transplanting culture of plants have an effect of enhancing the cold resistance of the stylosanthes guianensis.
In summary, the breeding and propagation method of the invention for the cold-resistant stylosanthes guianensis can obtain the cold-resistant stylosanthes guianensis with high cold resistance, high yield and high quality by combining sexual propagation and asexual propagation, and has great significance for expanding the application range and the application field thereof.
Claims (9)
1. A method for breeding and propagating cold-resistant stylosanthes guianensis is characterized by comprising the following steps:
1) selecting Oxley stylosanthes guianensis as a female parent and stylosanthes guianensis as a male parent;
2) seed pretreatment: carrying out low-frequency current treatment on Oxley stylosanthes guianensis seeds, and soaking the Oxley stylosanthes guianensis seeds in Streptomyces meliloti liquid;
soaking the stylosanthes guianensis seeds in a mixed solution of trichoderma reesei, humus and an abscisic acid aqueous solution, and alternately performing ice bath and hot water bath;
3) and (3) treating planting soil: red sandy soil, pH 5-6, organic matter content 1-1.5%, total nitrogen content 0.02-0.05%, total phosphorus content 0.05-0.08%, total potassium content 0.4-0.6%, without irrigation;
4) carrying out field planting on the pretreated Oxley stylosanthes guianensis seeds and the stylosanthes guianensis seeds, selecting strong plants, and spraying a sucrose and boric acid aqueous solution on stylosanthes guianensis inflorescences in the bud period;
performing artificial emasculation on Oxley stylosanthes guianensis in the bud stage, performing artificial pollination on the pollen of the stylosanthes guianensis in the bud stage, performing hybridization to obtain F1 generations, and selecting the seedling with better growth vigor as the cold-resistant stylosanthes guianensis seedling;
5) transplanting selected cold-resistant stylosanthes guianensis seedlings, 1-2 seedlings are transplanted in each hole, and uniformly spraying growth bacterium liquid on the leaves of the seedlings for natural growth;
6) and when branches grow out from the cold-resistant stylobactrium roseum seedlings, selecting the branches as seedlings, and carrying out cultivation and planting to obtain the cold-resistant stylobactrium roseum plants.
2. The method for breeding and propagating Styrax hancei as claimed in claim 1, wherein the concentration of the Streptomyces mixotrophicus in step 2) is 0.3-0.5%, and the Streptomyces mixus is a mixed solution of Streptomyces mixicola powder and deionized water.
3. A breeding and propagating method of han zhu Huacao as claimed in claim 1, wherein the solid-to-liquid ratio in the mixed solution of trichoderma reesei, humus and abscisic acid aqueous solution in step 2) is (3-5): (15-20) g/mL.
4. The method for breeding and propagating the winter jasmine flower and grass as claimed in claim 4, wherein the mass ratio of the trichoderma reesei to the humus is 1: (70-85);
the concentration of the abscisic acid aqueous solution is 25-35%.
5. A breeding method of han-kan stylosanthes guianensis as claimed in claim 1, wherein the low-frequency current treatment in step 2) is carried out for 8-12 min at 40-50 Hz for 3-4 times at intervals of 20-30 min.
6. The method for breeding and propagating Styrax hancei as claimed in claim 1, wherein the growth bacterial liquid in step 3) is a mixed bacterial liquid of Geospora and Trichoderma harzianum T-22, the concentration is 0.3-0.5%, and the mass ratio of the bacterial powders of Geospora and Trichoderma harzianum T-22 is (1-2): (2-5).
7. A breeding method of han zhu hua cao as claimed in claim 1, wherein in the aqueous solution of sucrose and boric acid in step 4), the concentration of sucrose is 10-20% and the concentration of boric acid is 0.01-0.02%.
8. The method for breeding and propagating the Stylosanthes guianensis according to claim 1, wherein the specific operation in step 5) is: selecting branches with the length of 30-35 cm as seedlings, planting in 3-ten days of the month, soaking grass seedlings into roots for 12-24 hours before planting the seedlings, burying soil with the depth of 10-15 cm, vertically transplanting, pouring root fixing water, and then performing field management and disease control.
9. The method for selectively breeding cold-resistant stylosanthes guianensis as claimed in claim 1, wherein the areas for crossing in step 4), transplanting in step 5) and cultivating in step 6) are stylosanthes guianensis pastures in south China.
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