阅读说明：本技术 一种利用化学诱变剂培育蔷薇变异新品种的方法 (Method for cultivating new rose variant variety by using chemical mutagen ) 是由 李承秀 于永畅 程甜甜 孙忠奎 张�林 燕语 张安琪 任红剑 杜辉 仲凤维 鹿苹 于 2021-08-14 设计创作，主要内容包括：本发明公开了一种利用化学诱变剂培育蔷薇变异新品种的方法,具体为：将蔷薇种子经沙藏出芽后播种在穴盘内,将平阳霉素和磷酸盐缓冲液混合制备平阳霉素溶液,将平阳霉素和MS液体培养基混合制备MS营养液；幼苗出土,待子叶展平真叶萌动时,将幼苗放入MS营养液中进行水培处理,采用平阳霉素溶液处理幼苗茎尖生长点,每天早、中、晚各处理一次,连续处理2～4天,处理完成后用清水冲洗幼苗茎尖生长点和根部,并转移到MS液体培养基中培养；处理一个月后,蔷薇的叶片逐渐出现变异,变异稳定后,将有变异性状的植株在温室内进行常规管理。该方法采用化学诱变剂处理植株茎尖生长点和根尖,操作简单,植株诱变率高,诱变效果好,植株的成活率高。(The invention discloses a method for cultivating a new rose variant variety by using a chemical mutagen, which comprises the following steps: sowing rose seeds in a plug tray after the rose seeds are subjected to sand storage and budding, mixing pingyangmycin and a phosphate buffer solution to prepare pingyangmycin solution, and mixing pingyangmycin and an MS liquid culture medium to prepare an MS nutrient solution; the method comprises the following steps of (1) enabling seedlings to emerge, placing the seedlings into MS nutrient solution for water culture treatment when cotyledons are flattened and true leaves germinate, treating stem tip growing points of the seedlings by using a pingyangmycin solution once in the morning, in the middle and at night every day, continuously treating for 2-4 days, flushing the stem tip growing points and roots of the seedlings by using clear water after the treatment is finished, and transferring the seedlings into an MS liquid culture medium for culture; after one month of treatment, the leaves of the rose gradually generate variation, and after the variation is stable, the plants with the variation characters are subjected to conventional management in a greenhouse. The method adopts chemical mutagen to treat the stem tip growing point and the root tip of the plant, and has the advantages of simple operation, high plant mutagenesis rate, good mutagenesis effect and high plant survival rate.)

1. A method for cultivating a new rose variant by using a chemical mutagen is characterized by comprising the following steps:

(1) collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare MS nutrient solution, wherein the concentration of pingyangmycin in pingyangmycin solution and the concentration of pingyangmycin in MS nutrient solution are both 55-65 mu g/ml; the method comprises the following steps of (1) enabling the rose seedling to come out of the soil in the last ten days of 4 months, placing the rose seedling into the prepared MS nutrient solution for water culture treatment when the cotyledon is flattened and the true leaf is exposed, treating the growth point of the stem tip of the rose seedling by using the prepared pingyangmycin solution, treating the growth point once in the morning, in the middle and at night every day for 2-4 days continuously, flushing the growth point and the root of the stem tip of the rose seedling by using clear water after the treatment is finished, and transferring the rose seedling to an MS liquid culture medium without the pingyangmycin for culture;

(3) after one month of treatment, the leaves of the rose gradually change, the color of the leaves begins to turn yellow, the change is stable from 9 months, the leaves become small and thick, the color of the leaves turns yellow or the leaves with speckles are formed, plants with the variation characters are managed in a greenhouse conventionally, and nutrient solution is sprayed for supplementing nutrition.

2. The method of claim 1, wherein the method comprises the steps of: in the step (1), the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler in a mass ratio of 4-6: 3-5: 2-4.

3. The method of claim 2, wherein the method comprises the steps of: the preparation method of the functional filler comprises the following steps: mixing and stirring the cellulose nanofiber suspension and a urea solution, and drying the obtained suspension by a spray drying method to obtain a cellulose nanofiber encapsulated urea material; mixing and stirring starch, glycerol and water uniformly, then drying to obtain thermoplastic starch, mixing the thermoplastic starch with a urea material encapsulated by poly (3-hydroxybutyrate) and cellulose nanofiber, adding the mixture into a torque rheometer, performing heat treatment for 5-15 min at 160-170 ℃ and 15-25 rpm, then performing hot pressing treatment for 6-8 min at 170-180 ℃ and 3-5 MPa, and finally cutting into particles to obtain the functional filler.

4. The method of claim 3, wherein the method comprises the steps of: the concentrations of the cellulose nanofiber suspension and the urea solution are respectively 1.5-2.5 wt% and 35-45 wt%, and the volume ratio of the cellulose nanofiber suspension to the urea solution is 1: 1; in the case of spray drying, the drying temperature was 180 ℃.

5. The method of claim 3, wherein the method comprises the steps of: the mass ratio of the starch, the glycerol, the poly (3-hydroxybutyrate) and the urea material encapsulated by the cellulose nano-fibers is 70: (25-35): 80: (50-60).

6. The method of claim 1, wherein the method comprises the steps of: in the step (2), the concentrations of the pingyangmycin in the pingyangmycin solution and the MS nutrient solution are both 60 mu g/ml.

7. The method of claim 1, wherein the method comprises the steps of: in the step (2), when the growing points of the stem tips of the rose seedlings are treated, absorbent cotton balls dipped with the pingyangmycin solution are placed on the growing points of the stem tips of the rose seedlings, and each treatment time is 8-12 min.

8. The method of claim 1, wherein the method comprises the steps of: in the step (3), the nutrient solution is a mixed solution of urea and monopotassium phosphate, and the mass concentrations of the urea and the monopotassium phosphate in the mixed solution are respectively 0.45-0.55% and 0.15-0.25%.

9. The method of claim 1, wherein the method comprises the steps of: in the step (3), the spraying frequency of the nutrient solution is once every 15 days.

Technical Field

The invention relates to the technical field of agriculture, in particular to a method for cultivating a new rose variant variety by using a chemical mutagen.

Background

The rose is a generic term for part of plants in the genus of roses, and mainly refers to the variety and horticultural species of rosa multiflora. Most of the flowers are rattan-shaped hedge flowers, and the stems are large and generally provided with hooks; leaf regeneration, odd feathers and compound leaves, 5-9 small leaves, teeth at the edge of the leaf, flat leaves but soft hair; the flowers are usually 6-7 florists, are coniform umbrella chamber inflorescences, grow on the tops of branches, have the diameter of about 3 cm, are opened once every year, and are important garden ornamental plants. The roses are cold resistant and can be used for medicine, and rosebush porridge prepared from the roses has good nutritional value. The systematic research on rose tree species resources is not carried out in China so far, the rose tree species resources are still in the conventional breeding stage of a lower utilization level, the artificial induced mutant is one of important ways for obtaining new plant germplasm, but the research on obtaining the rose tree species resources by artificially inducing the mutant is few at present, the variation of roses is low by using a common induction method, and the plant death rate is high in the variation process.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: aiming at the defects in the prior art, the method for cultivating the new rose variant variety by using the chemical mutagen is provided, the chemical mutagen is used for treating the stem tip growing point and the root tip of a plant, the operation is simple, the plant mutagenesis rate is high, the mutagenesis effect is good, and the self-made functional filler is added into the seedling culture substrate, so that the water and fertilizer retention performance of the seedling culture substrate is improved, and the survival rate of the plant is effectively improved.

In order to solve the technical problems, the technical scheme of the invention is as follows:

a method for cultivating a new rose variant by using a chemical mutagen comprises the following steps:

(1) collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare MS nutrient solution, wherein the concentration of pingyangmycin in pingyangmycin solution and the concentration of pingyangmycin in MS nutrient solution are both 55-65 mu g/ml; the rose seedling is put into the prepared MS nutrient solution for water culture treatment when the cotyledon is flattened and the true leaf is exposed, the prepared pingyangmycin solution is adopted to treat the growth point of the stem tip of the rose seedling, the treatment is carried out once in the morning, in the middle and at night every day, the treatment is continuously carried out for 2-4 days, the growth point and the root of the stem tip of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after one month of treatment, the leaves of the rose gradually change, the color of the leaves begins to turn yellow, the change is stable from 9 months, the leaves become small and thick, the color of the leaves turns yellow or the leaves with speckles are formed, plants with the variation characters are managed in a greenhouse conventionally, and nutrient solution is sprayed for supplementing nutrition.

Preferably, in the step (1), the seedling raising substrate is prepared by mixing turfy soil, sawdust and functional filler in a mass ratio of 4-6: 3-5: 2-4.

Preferably, in the above technical solution, the preparation method of the functional filler comprises: mixing and stirring the cellulose nanofiber suspension and a urea solution, and drying the obtained suspension by a spray drying method to obtain a cellulose nanofiber encapsulated urea material; mixing and stirring starch, glycerol and water uniformly, then drying to obtain thermoplastic starch, mixing the thermoplastic starch with a urea material encapsulated by poly (3-hydroxybutyrate) and cellulose nanofiber, adding the mixture into a torque rheometer, performing heat treatment for 5-15 min at 160-170 ℃ and 15-25 rpm, then performing hot pressing treatment for 6-8 min at 170-180 ℃ and 3-5 MPa, and finally cutting into particles to obtain the functional filler.

Preferably, the concentrations of the cellulose nanofiber suspension and the urea solution are 1.5-2.5 wt% and 35-45 wt%, respectively, and the volume ratio of the cellulose nanofiber suspension to the urea solution is 1: 1; in the case of spray drying, the drying temperature was 180 ℃.

Preferably, in the above technical solution, the mass ratio of the starch, the glycerol, the poly (3-hydroxybutyrate), and the urea material encapsulated by the cellulose nanofiber is 70: (25-35): 80: (50-60).

Preferably, in the step (2), the concentration of pingyangmycin in the pingyangmycin solution and the concentration of pingyangmycin in the MS nutrient solution are both 60 mug/ml.

Preferably, in the step (2), the absorbent cotton ball dipped with the pingyangmycin solution is placed on the growing point of the stem tip of the rose seedling, and the treatment time is 8-12 min each time.

Preferably, in the step (3), the nutrient solution is a mixed solution of urea and monopotassium phosphate, and the mass concentrations of urea and monopotassium phosphate in the mixed solution are 0.45-0.55% and 0.15-0.25%, respectively.

Preferably, in the step (3), the nutrient solution is sprayed once every 15 days.

Due to the adoption of the technical scheme, the invention has the beneficial effects that:

the method comprises the steps of mixing pingyangmycin and a phosphate buffer solution to prepare pingyangmycin solution, mixing pingyangmycin and an MS liquid culture medium to prepare MS nutrient solution, transferring unearthed seedlings into the MS nutrient solution, putting absorbent cotton balls dipped with the pingyangmycin solution on stem tip growing points of the seedlings, and treating the stem tip growing points and root tips of rose seedlings, so that the variation rate of rose plants is greatly improved, and the variation effect is good.

The functional filler is prepared by encapsulating urea by adopting cellulose nano-fiber and then coating thermoplastic starch and poly (3-hydroxybutyrate), and has good water and fertilizer retention performance; the poly (3-hydroxybutyrate) is biodegraded in an aerobic and anaerobic environment without forming toxic waste; according to the invention, glycerin is used as a plasticizer to prepare thermoplastic starch, and the thermoplastic starch is blended with poly (3-hydroxybutyrate) to coat a urea material encapsulated by cellulose nano-fibers, so that the prepared functional filler has excellent performance, can effectively provide nutrition required by rose seedlings during mutagenesis, and greatly reduces the death rate of plants. The mutagenesis method provided by the invention is simple to operate, high in plant mutagenesis rate, good in mutagenesis effect and low in plant mortality.

Detailed Description

The invention is further illustrated by the following examples. It should be understood that these examples are for illustrative purposes only and are not intended to limit the scope of the present invention.

The following examples and comparative examples were prepared with the functional filler:

mixing and stirring 100ml of cellulose nanofiber suspension with the concentration of 1.5-2.5 wt% and 100ml of urea solution with the concentration of 35-45 wt%, and drying the obtained suspension by a spray drying method to obtain a cellulose nanofiber encapsulated urea material; mixing 70g of starch, 30g of glycerol and 20g of water, uniformly stirring, drying to obtain thermoplastic starch, mixing 70g of thermoplastic starch with 70g of poly (3-hydroxybutyrate) and 50g of urea material encapsulated by cellulose nanofiber, adding the mixture into a torque rheometer, performing heat treatment at 165 ℃ and 20rpm for 10min, performing hot pressing treatment at 175 ℃ and 5MPa for 7min, and finally cutting into granules to obtain the functional filler.

Example 1

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:3: 2;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out for 2 days continuously, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Example 2

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 6:5: 2;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Example 3

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 6:4: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Example 4

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 5:4: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when the cotyledon flattening true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on the stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out for 2 days continuously, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Example 5

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when the cotyledon flattening true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on the stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is continuously carried out for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 1

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil and wood chips according to the mass ratio of 7: 5;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 60 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when the cotyledon flattening true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on the stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is continuously carried out for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 2

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a plug tray filled with a seedling substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) the seedlings are unearthed in the last ten days of 4 months, when cotyledons are flattened and true leaves are exposed, the rose seedlings are put into an MS liquid culture medium for water culture treatment, and nutrient solution consisting of urea and potassium dihydrogen phosphate is sprayed to supplement nutrition and is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 3

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin and an MS liquid culture medium to prepare an MS nutrient solution; the concentration of pingyangmycin in the MS nutrient solution is 60 mug/ml; the seedlings of the rose come out of the soil in the last ten days of 4 months, when cotyledons are flattened and true leaves are exposed, the rose seedlings are put into the prepared MS nutrient solution for water culture treatment for 3 days continuously, the roots of the rose seedlings are washed by clear water after the treatment is finished, and then the rose seedlings are transferred to an MS liquid culture medium without adding pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 4

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 10 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 5

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 30 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 6

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 50 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 7

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 70 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 8

(1) Collecting rose seeds from the bottom of 10 months to the beginning of 11 months, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate after the rose seeds are subjected to sand storage and sprouting, and performing conventional management after the rose seeds emerge; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) mixing pingyangmycin with a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, and mixing pingyangmycin with an MS liquid culture medium to prepare an MS nutrient solution; the concentrations of pingyangmycin in pingyangmycin solution and MS nutrient solution are both 80 mug/ml; 4, the rose seedling is unearthed in the last ten days of the month, when cotyledons are flattened and true leaves are exposed, the rose seedling is placed into the prepared MS nutrient solution for water culture treatment, the absorbent cotton ball dipped with the pingyangmycin solution is placed on a stem tip growing point of the rose seedling for treatment, the treatment is carried out once in the morning, in the middle and at the evening every day, the treatment is carried out for 10min every time, the treatment is carried out continuously for 3 days, the stem tip growing point and the root of the rose seedling are washed by clear water after the treatment is finished, and then the rose seedling is transferred to an MS liquid culture medium without the pingyangmycin for culture;

(3) after the rose leaves are treated for one month, the rose leaves gradually generate variation, the variation of the rose leaves is obvious when the variation is stable in 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and monopotassium phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

Comparative example 9

(1) Mixing pingyangmycin and a phosphate buffer solution with the pH value of 7 to prepare a pingyangmycin solution, soaking rose seeds collected from the bottom of 10 months to the beginning of 11 months in the pingyangmycin solution for 3 days, taking out the rose seeds, cleaning the rose seeds, storing the rose seeds in sand, sprouting the rose seeds, sowing the rose seeds in a seedling raising tray filled with a seedling raising substrate, and performing conventional management after sprouting; the seedling raising substrate is formed by mixing turfy soil, sawdust and functional filler according to the mass ratio of 4:5: 3;

(2) the seedlings come out of the earth in the last ten days of 4 months, and when cotyledons are flattened and true leaves sprout, the rose seedlings are cultured in an MS liquid culture medium without adding the pingyangmycin;

(3) after one month, the leaves of the rose gradually generate variation, the variation of the leaves is stable until 9 months, the variation plants mainly show that the leaves become small and thick, the leaf color becomes yellow or the leaves with speckles are formed, the plants with the variation characters are managed conventionally in a greenhouse, nutrient solution consisting of urea and potassium dihydrogen phosphate is sprayed for supplementing the nutrition, and the nutrient solution is sprayed once every 15 days; the mass concentrations of urea and potassium dihydrogen phosphate in the nutrient solution are 0.2% and 0.2%, respectively.

The total number of the plants treated is 300, and the death number and the variation number of the plants are counted after the variation of the plants is stable, and the results are shown in table 1.

TABLE 1

From the results, the plant mutation rate is increased along with the increase of the pingyangmycin concentration, the mutation rate of the plant is reduced when the pingyangmycin is increased to a certain concentration and then is increased continuously, the concentration of the pingyangmycin is controlled at 60 mu g/ml, and the mutation rate of the plant is the highest. Compared with the method that the plant root tip is only treated by adopting the pingyangmycin, the variation rate of the plant is higher when the root tip and the growing point of the stem tip are simultaneously treated. In addition, compared with the method adopting the pingyangmycin solution to treat seeds for mutagenesis, the mutation rate of the plants is higher when stem tip growing points and root tips of the plants are subjected to mutagenesis treatment. According to the invention, a certain functional filler is added into the seedling culture substrate, so that the survival rate of plants is improved on the premise of not influencing the variation rate of the plants.

Further, it should be understood that various changes or modifications of the present invention may be made by those skilled in the art after reading the teaching of the present invention, and such equivalents may fall within the scope of the present invention as defined in the appended claims.