CN115266617A - Screening method of low-temperature-resistant garden chrysanthemum varieties in plateau area - Google Patents
Screening method of low-temperature-resistant garden chrysanthemum varieties in plateau area Download PDFInfo
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Abstract
The invention discloses a method for screening low-temperature-resistant garden chrysanthemum varieties, which can be used for evaluating the low-temperature adaptability of garden chrysanthemum through the determination of 2 root indexes and 1 photosynthetic pigment index of the garden chrysanthemum, so as to screen the low-temperature-resistant garden chrysanthemum varieties. The method can identify the low-temperature-resistant varieties of the garden chrysanthemum by only measuring 3 indexes, and has the advantages of low identification cost, simple operation and easy popularization.
Description
Technical Field
The invention relates to a method for screening garden chrysanthemum, in particular to a method for screening low-temperature-resistant garden chrysanthemum varieties in a plateau area, and belongs to the technical field of chrysanthemum variety screening.
Background
Chrysanthemum morifolium (Chrysanthemum morifolium) is a perennial root herbaceous plant of the Chrysanthemum genus of the Compositae family, wherein the garden Chrysanthemum has the characteristics of short and compact plant type, multiple varieties and strong ornamental value, becomes a good ground cover material for urban greening, and asexual cuttage propagation is adopted in production, so that the Chrysanthemum plays an important role in the flower market due to fast growth and high propagation coefficient.
In recent years, with environmental construction and new variety breeding, a plurality of chrysanthemum varieties are introduced in various regions, but the requirement on temperature in the regrowth process of chrysanthemum is strict, the temperature is one of key factors influencing cuttage, the germination and growth of new roots are not facilitated due to overhigh or overlow temperature, and even the chrysanthemum cuttage seedlings have the phenomena of slow rooting, poor development of root systems and cutting slips and the like due to low temperature (Tsipouridis et al, 2005; li Xiaowei and the like, 2014).
The Qinghai area where the inventor is located also introduces a plurality of chrysanthemum varieties. However, the growth and development of the cutting seedlings are seriously influenced due to high altitude and low temperature at the Qinghai-Tibet plateau, the survival rate of the cutting seedlings is greatly reduced, and the problems of long rooting time of the cutting seedlings, poor root system quality, poor growth and development of cutting slips and the like are caused. Although most of production adopts sunlight greenhouses, the overall foundation is weak, the illumination, temperature and controllability are poor, the seedling raising cost of a heating system is high, the low temperature in spring becomes a restriction factor of chrysanthemum facility cultivation in plateau areas, and the time for cutting and rooting is longer than that in other areas. Therefore, when chrysanthemum varieties are introduced in Qinghai areas, rooting seedlings are introduced, and the application of chrysanthemum in Qinghai areas is seriously influenced due to high price and large volume of the rooting seedlings. Researches on low-temperature-resistant variety screening and adaptability evaluation of the garden chrysanthemum are not reported, and blindness is caused when the chrysanthemum varieties are introduced in the Qinghai region.
Disclosure of Invention
Aiming at the defects of the prior art, the invention aims to provide a simple and practical screening method for low-temperature-resistant varieties of chrysanthemum morifolium.
The invention relates to a screening method of low-temperature-resistant garden chrysanthemum varieties in plateau areas, which comprises the following steps:
(1) Cutting garden chrysanthemum cuttings, cutting in a plug tray, and setting 2 temperatures for cutting treatment: the stress group is at 15 ℃/15 ℃ day/night and the control group is at 25 ℃/20 ℃;
(2) Determining the total surface area and the number of root tips of the cutting shoots of the garden chrysanthemum: during the growth period of the chrysanthemum morifolium ramat, the total surface area of roots and the number of root tips are measured in cm respectively2And root as a unit;
(3) Determining the total chlorophyll content of the cutting slips of the garden chrysanthemum: extracting chlorophyll from fresh garden chrysanthemum samples to be detected by using ethanol, pouring the chlorophyll extracting solution into a cuvette, taking an ethanol solution as a control, carrying out zero setting, and measuring the absorbance under 665nm and 649nm wavelengths; calculated according to a formulaChlorophyll a, chlorophyll b and total chlorophyll concentration to obtain total chlorophyll content in the tissue; wherein: chlorophyll a concentration =13.95A665-6.88A649(ii) a Chlorophyll b concentration =24.96A649-7.32A665(ii) a Total chlorophyll concentration = chlorophyll a concentration + chlorophyll b concentration; chloroplast pigment content = (pigment concentration x extract volume)/sample fresh weight;
(4) Calculating the low temperature resistance coefficient of the index measured in the step (2) and the step (3): root total surface area low temperature resistance coefficient X1= 100% (post-stress assay/control assay); total chlorophyll content low temperature resistant coefficient X2= 100% (post-stress assay/control assay); root number low temperature resistance coefficient X3= 100% (post-stress assay/control assay);
(5) Calculating comprehensive evaluation value =0.220+0.090X of low temperature resistance1+0.580X2+0.095X3(ii) a The higher the comprehensive evaluation value score of low temperature resistance, and X1、X2And X3When all the varieties are not zero, the varieties are more low-temperature resistant varieties.
The cutting of the garden chrysanthemum in the step (1) can be cut according to a conventional method in the field, in a specific example, 8-10 cm of top tips of the garden chrysanthemum are cut to serve as the cutting, 2-3 leaves on the upper portion are reserved, and the cutting is carried out in a plug tray.
The growth period of the garden chrysanthemum in the step (2) refers to 26-28 days after cuttage.
The total surface area and the number of the root tips of the cutting roots of the garden chrysanthemum in the step (2) can be measured according to a root scanning system commonly used in the field, such as a root phenotype analysis system GXY-A, root phenotype analysis software Win-RHIO and other root scanning analysis software.
The total chlorophyll content of the cutting slips of the garden chrysanthemum in the step (3) can be determined according to a conventional method in the field, and in order to enable the determination result to be more uniform and accurate, the application provides a specific determination method which comprises the following steps: respectively taking fresh garden chrysanthemum samples to be detected of a stress group and a control group, cleaning surface dirt, shearing, uniformly mixing, respectively weighing 0.1g of sheared fresh samples, and respectively putting 3 parts of sheared fresh samples into 10mL centrifuge tubes; adding 5mL of 95% ethanol into each centrifugal tube, and completely soaking the sample in the ethanol solution; soaking in dark for 24 hr, pouring chlorophyll extractive solution into a cuvette, adjusting to zero with 95% ethanol solution as control, and measuring absorbance at 665nm and 649 nm.
The higher the score of the comprehensive evaluation value of the low-temperature resistance in the step (5) of the invention is, the more low-temperature resistant varieties are, and when the score is more than or equal to 0.85, the varieties of the low-temperature resistant garden chrysanthemum are, namely, the varieties which can grow by cuttage at the temperature of 10-25 ℃.
In some embodiments, the garden chrysanthemum is jinling yulan, jinling honghe, jinling fireworks, jinling yanyan, jinling poinsettia or jinling billow.
Compared with the prior art, the invention has the beneficial effects that:
the invention provides a simple, convenient and effective method for screening low-temperature-resistant varieties of garden chrysanthemum in plateau areas, and the low-temperature adaptability of the garden chrysanthemum can be evaluated by measuring 2 root indexes and 1 photosynthetic pigment index of the garden chrysanthemum, so that the low-temperature-resistant varieties of the garden chrysanthemum can be screened. The method can identify the low-temperature-resistant varieties of the garden chrysanthemum by only measuring 3 indexes, and has the advantages of low identification cost, simple operation and easy popularization.
The method establishes a reliable comprehensive evaluation method, can effectively screen out varieties with low temperature in the cuttage-resistant period, and has important significance for introduction and industry of the small chrysanthemum gardens in the plateau areas.
Detailed Description
The following examples are given to facilitate a better understanding of the invention, but do not limit the invention. The experimental procedures in the following examples are conventional unless otherwise specified. The test materials used in the following examples were purchased from a conventional biochemical reagent store unless otherwise specified.
Unless otherwise specified, the experimental materials of the following examples are as follows: 6 garden chrysanthemum varieties introduced by Nanjing agriculture university are used as test materials, namely Jinling billows (pink), jinling yan (purple), jinling poinsettia (red), jinling fireworks (compound color), jin Lingfen magnolia (pink purple) and Jinling red lotus (red). The flowering season is the intersection between the middle 8 th-9 th-month late and the travel season of Qinghai. 6 cutting seedlings of garden chrysanthemum varieties are planted in a greenhouse of a gardening institute of Qinghai university, a random block is adopted, the plant-row spacing is 40 multiplied by 40cm, and the same conventional water and fertilizer management method is adopted.
Example 1
The invention screens the introduced 6 garden chrysanthemum varieties of Nanjing agriculture university for low temperature resistant varieties, and the specific test materials are shown in Table 1.
TABLE 1 Garden chrysanthemum variety information sheet
The test materials were planted in the test field of the garden institute of academy of agriculture and forestry, qinghai province, 5 months in 2021, the planting spacing was 40cm, the row spacing was 40cm, the materials were randomly arranged, and the same conventional water and fertilizer management method was adopted. In the test, a strong and disease and pest free stock plant is selected, 8-10 cm of top shoots are cut to be used as cutting slips, 2-3 leaves at the upper part are reserved and are cut in a 50-hole plug tray, and 2 temperature treatments are set in total: the material was subjected to different temperature conditions at 15 ℃/15 ℃ and 25 ℃/20 ℃ day/night, respectively, with 25 ℃ as a control, and 50 cuttings were treated each.
Screening of low-temperature-resistant garden chrysanthemum varieties comprises the following steps:
(1) Determination of the total surface area and the number of root tips of the garden chrysanthemum roots:
during the growth period of the garden chrysanthemum morifolium, the total surface area of roots and the number of root tips are measured by a root system phenotype analysis system GXY-A, and the total surface area of the roots is expressed in cm2The number of root tips is in units of roots.
(2) Measuring the total chlorophyll content of the garden chrysanthemum: in the growth period of the garden chrysanthemum, the total chlorophyll content in leaves is measured: selecting top tips of various varieties under different temperature treatment, cleaning dirt on the surfaces of the top tips, shearing, uniformly mixing, weighing 0.1g of sheared fresh samples and 3 parts in total, and respectively placing the weighed sheared fresh samples into 10mL centrifuge tubes. 5mL of 95% ethanol was added to each centrifuge tube, and the samples were completely soaked in the ethanol solution.Soaking in dark for 24 hr, pouring chlorophyll extractive solution into a cuvette, adjusting to zero with 95% ethanol solution as control, and measuring absorbance at 665nm and 649 nm. Calculating the chlorophyll a, chlorophyll b and total chlorophyll concentration according to a formula so as to obtain the content of the pigment in the tissue. (chlorophyll a concentration = 13.95A)665-6.88A649(ii) a Chlorophyll b concentration =24.96A649-7.32A665(ii) a Total chlorophyll concentration = chlorophyll a concentration + chlorophyll b concentration; chloroplast pigment content = (pigment concentration x volume of extract)/fresh weight of sample)
(3) Calculating the low temperature resistance coefficient alpha of the indexes measured in the step (1) and the step (2): root total surface area low temperature resistance coefficient X1= 100% (post-stress assay/control assay); total chlorophyll content low temperature resistant coefficient X2= 100% (post-stress assay/control assay); root number low temperature resistance coefficient X3= 100% (post stress assay/control assay).
(4) And (3) evaluating the low-temperature adaptability of the garden chrysanthemum: the method comprises the following steps of carrying out low temperature adaptability evaluation through an optimal regression equation established by taking the low temperature resistance coefficient of each single index of the garden chrysanthemum as an independent variable, and calculating according to the following formula:
comprehensive evaluation value of low temperature resistance =0.220+0.090X1+0.580X2+0.095X3。
The screening results of the low-temperature resistance of the garden chrysanthemum are shown in table 2, the higher the comprehensive evaluation value score (D value) of the low-temperature resistance is, the stronger the low-temperature resistance of the material is, wherein the maximum D value of the Jinling billow ' indicates that the variety has the strongest low-temperature resistance, and the score ordering is proved to be consistent with the actual low-temperature resistance (Sun Donglei and the like, 2021), wherein the Jinling billow ' and the Jinling red load ' are in the same category, belong to the high low-temperature resistant type and are varieties capable of growing by cuttage at the temperature of 10-25 ℃.
TABLE 2 comprehensive evaluation results of low temperature resistance of chrysanthemum morifolium
The inventor of the application also researches the morphological difference of 6 garden chrysanthemum under different temperatures, and further verifies the reliability of the scheme of the application.
The six species of chrysanthemum in the field of the embodiment 1 are respectively selected from mother plants which grow robustly and have no plant diseases and insect pests, 8-10 cm of top tips are cut to be used as cutting slips, 2-3 leaves at the upper part are reserved and are cut in a 50-hole plug tray, and 4 temperature treatments are set in total: 5 deg.C, 10 deg.C, 15 deg.C, 25 deg.C. And (3) respectively placing the materials at different temperatures, processing 50 cutting slips for each time, respectively sampling 28 days after cutting, and observing the rooting and development conditions.
The result shows that the roots of all varieties can normally grow under normal culture (25 ℃); all varieties except 'Jinling fireworks display' take root under the treatment of 15 ℃; at 10 ℃, only 'Jinling red lotus' and 'Jinling waves' have root systems generated, and the rest varieties do not root; this is also consistent with the results of the screening described in example 1, further demonstrating the reliability of the method described in this application.
Claims (9)
1. A screening method for low-temperature-resistant garden chrysanthemum varieties in plateau areas is characterized by comprising the following steps:
(1) Cutting garden chrysanthemum cuttings, cutting in a plug tray, and setting 2 temperatures for cutting treatment: the stress group is at 15 ℃/15 ℃ day/night and the control group is at 25 ℃/20 ℃;
(2) Determining the total surface area of the root and the number of root tips of the cutting shoot of the garden chrysanthemum: during the growth period of the chrysanthemum morifolium ramat, the total surface area of roots and the number of root tips are measured in cm respectively2And root as a unit;
(3) Determining the total chlorophyll content of the cutting slips of the garden chrysanthemum: extracting chlorophyll from fresh garden chrysanthemum samples to be detected by using ethanol, pouring the chlorophyll extracting solution into a cuvette, using an ethanol solution as a control, carrying out zero setting, and measuring the absorbance under the wavelengths of 665nm and 649 nm; calculating the chlorophyll a, chlorophyll b and total chlorophyll concentration according to a formula to obtain the total chlorophyll content in the tissue; wherein: chlorophyll aConcentration =13.95A665-6.88A649(ii) a Chlorophyll b concentration =24.96A649-7.32A665(ii) a Total chlorophyll concentration = chlorophyll a concentration + chlorophyll b concentration; chloroplast pigment content = (pigment concentration x extract volume)/sample fresh weight;
(4) Calculating the low temperature resistance coefficient of the index measured in the step (2) and the step (3): root total surface area low temperature resistance coefficient X1= 100% (post-stress assay/control assay); total chlorophyll content low temperature resistant coefficient X2= 100% (post-stress assay/control assay); root number low temperature resistance coefficient X3= 100% (post-stress assay/control assay);
(5) Calculating comprehensive evaluation value =0.220+0.090X of low temperature resistance1+0.580X2+0.095X3(ii) a The higher the comprehensive evaluation value score of the low-temperature resistance, and X1、X2And X3When all the varieties are not zero, the varieties are more low-temperature resistant varieties.
2. The screening method of low temperature resistant garden chrysanthemum varieties according to claim 1, wherein 8-10 cm of top tips of garden chrysanthemum are cut in the step (1) to serve as cutting slips, 2-3 leaves on the upper portion are reserved, and the cutting slips are placed in a plug tray.
3. The screening method of low temperature resistant garden chrysanthemum varieties according to claim 1, wherein the growth period of garden chrysanthemum in step (2) is 26-28 days after cuttage.
4. The screening method of low temperature resistant garden chrysanthemum varieties as claimed in claim 1, wherein the total surface area of the cutting roots and the number of the root tips of the garden chrysanthemum in step (2) are measured by a root scanning system.
5. The screening method of low-temperature-resistant garden chrysanthemum varieties according to claim 4, wherein the root scanning system is a root phenotype analysis system GXY-A or root phenotype analysis software Win-RHIO.
6. The screening method of low-temperature-resistant garden chrysanthemum varieties according to claim 1, wherein the method for determining the total chlorophyll content of the garden chrysanthemum cuttings in the step (3) specifically comprises the following steps: respectively taking fresh garden chrysanthemum samples to be detected of a stress group and a control group, cleaning surface dirt, shearing, uniformly mixing, respectively weighing 0.1g of sheared fresh samples, and respectively putting 3 parts of sheared fresh samples into 10mL centrifuge tubes; adding 5mL of 95% ethanol into each centrifugal tube, and completely soaking the sample in the ethanol solution; soaking in dark for 24 hr, pouring chlorophyll extractive solution into a cuvette, adjusting to zero with 95% ethanol solution as control, and measuring absorbance at 665nm and 649 nm.
7. The screening method of low temperature resistant garden chrysanthemum varieties according to claim 1, wherein the low temperature resistant garden chrysanthemum varieties are selected when the score is more than or equal to 0.85.
8. The screening method of low temperature resistant garden chrysanthemum varieties according to claim 7, wherein the low temperature resistant garden chrysanthemum varieties are varieties capable of growing by cutting at a temperature of 10-25 ℃.
9. The screening method of low-temperature-resistant garden chrysanthemum varieties according to claim 1, wherein the garden chrysanthemum is jinling powdery yulan, jinling red lotus, jinling fireworks, jinling yanyan, jinling poinsettia or jinling waves.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116267454A (en) * | 2023-02-02 | 2023-06-23 | 安徽农业大学 | Method for screening cold-resistant varieties of non-heading Chinese cabbages |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN116267454A (en) * | 2023-02-02 | 2023-06-23 | 安徽农业大学 | Method for screening cold-resistant varieties of non-heading Chinese cabbages |
| CN116267454B (en) * | 2023-02-02 | 2023-11-24 | 安徽农业大学 | Method for screening cold-resistant varieties of non-heading Chinese cabbages |
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