CN111771702A - Method for evaluating saline-alkali resistance of industrial hemp in seedling stage by using water culture method - Google Patents
Method for evaluating saline-alkali resistance of industrial hemp in seedling stage by using water culture method Download PDFInfo
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- A—HUMAN NECESSITIES
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Abstract
A method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using a water culture method belongs to the technical field of analysis of the saline-alkali resistance of industrial hemp. In order to solve the problems that the existing identification of the saline-alkali property of the industrial hemp at the seedling stage consumes long time, occupies large space and the like, 3 layers of filter paper are respectively placed on the upper part and the lower part of the industrial hemp, soaked and cultured in the dark at the temperature of 20-25 ℃ for 2-3 days; preparing nutrient solution, diluting to 2/3 times, culturing in light-tight plastic box, placing the culture solution in the box, placing floating plate, perforating on the foam, and placing cotton and germinated seed in the holes; when 3 pairs of true leaves grow, dividing the seedlings into a control group and a treatment group; respectively collecting plant samples of different varieties as controls, and determining the agronomic characters; in cultureNaHCO is added into nutrient solution3The control group is normal culture solution; and (5) determining the agronomic characters for analysis. The method is completed in about 27 days, can scientifically and reasonably evaluate the salt and alkali tolerance of industrial hemp, and greatly shortens the evaluation period.
Description
Technical Field
The invention belongs to the technical field of analysis of saline-alkali resistance of industrial hemp, and particularly relates to a method for evaluating the saline-alkali resistance of industrial hemp in a seedling stage by using a hydroponic method.
Background
Industrial Cannabis sativa (Cannabis sativa L.) refers to a variety of Cannabis sativa having a Tetrahydrocannabinol (THC) content of less than 0.3%, and is an annual herb plant of the Cannabis genus (Cannabis) of the Cannabinaceae family (Cannabinaceae). The Chinese ancient book of agriculture "Ziming Yao shu" has been well documented for the technology of hemp planting. At the beginning of the establishment of new china, research on cannabis has mainly focused on high-yield cultivation experience. In recent years, in order to promote the development of industrial hemp industry, breeding experts breed a plurality of special industrial hemp varieties according to the uses such as fiber type, seed type and the like, such as: hemp series, Yunyan hemp series, Qingma series, Fenma series, etc.
The Heilongjiang province has 66.7 million hectares of saline-alkali soil, and has a trend of increasing year by year, so that the yield of grains and grasses is seriously influenced, and the development of farming and animal husbandry is further limited. With the supply side structural reform and the planting structural adjustment, Heilongjiang province gradually becomes the main planting province of industrial hemp. The industrial hemp has strong adaptability and stronger salt-alkali tolerance. The Chinese hemp industry technology system transfers hemp production to non-cultivated land (saline-alkali land, hilly land), but industrial hemp is not suitable for planting in all types of saline-alkali land, and the salt-alkali tolerance of industrial hemp varieties needs to be evaluated rapidly and scientifically so as to plant the industrial hemp scientifically and reasonably.
At present, in northeast China, industrial hemp can be planted for only one season every year, and the evaluation of the alkali resistance of the industrial hemp by field planting wastes resources. The time is long, at least 40 days are needed for completion, and the method is mainly completed through potting, so that the required space is large, and more resources are needed.
Disclosure of Invention
The invention aims to solve the problems of long time consumption, large occupied space and the like in the conventional identification of the saline-alkali property of the industrial hemp in the seedling stage, and provides a method for evaluating the saline-alkali resistance of the industrial hemp in the seedling stage by using a water culture method.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
a method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using a hydroponic method comprises the following steps:
the method comprises the following steps: respectively placing 3 layers of filter paper on the upper and lower parts of different varieties of industrial hemp, soaking the industrial hemp in distilled water, and culturing the industrial hemp in the dark at the temperature of 20-25 ℃ for 2-3 days;
step two: preparing a nutrient solution for water culture, wherein the nutrient solution comprises the following specific components: 945mg/L of calcium nitrate tetrahydrate, 404mg/L of potassium nitrate, 120mg/L of urea, 230mg/L of ammonium dihydrogen phosphate, 246mg/L of magnesium sulfate, 1ml/L of iron salt solution and 6.5 parts of trace element solution 2ml/L, pH;
step three: diluting the concentration of the nutrient solution in the second step to 2/3 times, culturing the industrial hemp obtained in the first step in a light-tight plastic box, filling the culture solution in the plastic box, putting the plastic box into a floating plate, punching the floating plate, wherein the distance between the holes is 2-5 cm, putting small cotton balls and the germinated seeds obtained in the first step into the holes, and putting the small cotton balls and the germinated seeds into an artificial climate box;
step four: evaluation of salt-base tolerance: when 3 pairs of true leaves grow, dividing the seedlings into a control group and a treatment group; respectively collecting plant samples of different varieties as controls, and determining the agronomic characters; adding NaHCO into the culture solution3Making the concentration of the culture medium reach 100mM, and the control group is normal culture medium; changing the culture solution every 2 days; and (5) surveying the number of dead seedlings as long as the day 5, collecting plant samples of different varieties, and determining the agronomic characters for analyzing the influence of saline-alkali stress on the growth of the industrial hemp.
Compared with the prior art, the invention has the beneficial effects that: the method utilizes a water culture method for identification, is completed within about 27 days, can carry out scientific and reasonable evaluation on the salt-base tolerance of the industrial hemp, and greatly shortens the evaluation period; meanwhile, the water culture method is utilized, so that the space and resources are saved. Through the evaluation and analysis of the salt-base tolerance of the germplasm resources, a resource basis is provided for the subsequent resistance breeding. Hydroponics (Hydroponics) is a novel soilless culture mode of plants, also called nutrient solution culture, and is characterized in that root systems of the plants are directly soaked in nutrient solution, and the nutrient solution can replace soil and provide growth factors such as water, nutrients, oxygen and the like for the plants, so that the plants can normally grow. After the hydroponics method is adopted, the plants can directly absorb nutrients and moisture in the nutrient solution through roots, and solutions with balanced nutrients can be manually prepared to realize balanced diet of the plants, which is obviously much easier than finding perfect soil.
Drawings
FIG. 1 is a schematic representation of the plants of Cannabis sativa I and golden knife-15 control group compared for 0 day;
FIG. 2 is a schematic representation of the plants of the Cannabis I and golden knife-15 control and treated groups growing for 5 days.
Detailed Description
The technical solutions of the present invention are further described below with reference to the drawings and the embodiments, but the present invention is not limited thereto, and modifications or equivalent substitutions may be made to the technical solutions of the present invention without departing from the spirit and scope of the technical solutions of the present invention.
The first embodiment is as follows: the embodiment describes a method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using a hydroponic method, which comprises the following steps:
the method comprises the following steps: respectively placing 3 layers of filter paper on the upper and lower parts of different varieties of industrial hemp, soaking the industrial hemp in distilled water, and culturing the industrial hemp in the dark at the temperature of 20-25 ℃ for 2-3 days; the above culturing process can be carried out in a culture dish, an incubator, a culture box or a germination box, for example, different varieties of industrial hemp are placed in the culture dish, 3 layers of filter paper are respectively placed on the upper part and the lower part of the industrial hemp, the industrial hemp is soaked in distilled water and placed in the incubator at the temperature of 25 ℃, the industrial hemp is cultured in the dark for 2 to 3 days, and the industrial hemp is transplanted into the culture box after the buds grow to about 1 cm; the germination speed is higher at 25 ℃.
Step two: preparing a nutrient solution for water culture, wherein the nutrient solution comprises the following specific components: 945mg/L of calcium nitrate tetrahydrate, 404mg/L of potassium nitrate, 120mg/L of urea, 230mg/L of ammonium dihydrogen phosphate, 246mg/L of magnesium sulfate, 1ml/L of iron salt solution and 6.5 parts of trace element solution 2ml/L, pH; according to the fertilizer requirement condition of industrial hemp, the Hoagland's nutrient solution is improved, so that various large quantities of trace elements such as nitrogen, phosphorus and potassium required by the growth of the industrial hemp can be provided, and the normal growth of the industrial hemp is ensured.
Step three: diluting the concentration of the nutrient solution in the second step to 2/3 times, culturing the industrial hemp obtained in the first step in a light-tight plastic box, filling the culture solution in the plastic box, wherein the amount of the culture solution needs to ensure that the root part is completely immersed, putting a floating plate (for example, plastic foam or other floatable objects) with the thickness of 1cm into the plastic box, punching holes on the floating plate, enabling the distance between every two holes to be 2-5 cm, putting small cotton balls and the germinated seeds obtained in the first step into the holes, and putting the small cotton balls and the germinated seeds into an artificial climate box;
step four: evaluation of salt-base tolerance: 18-22 days later, and waiting for the seedlingsGrowing to 3 pairs of true leaves, setting 20 seedlings in each box, and dividing into a control group and a treatment group; respectively collecting plant samples of different varieties as controls, and determining the agronomic characters; adding NaHCO into the culture solution3Making the concentration of the culture medium reach 100mM, and the control group is normal culture medium; changing the culture solution every 2 days; and (5) surveying the number of dead seedlings as long as the day 5, collecting plant samples of different varieties, and determining the agronomic characters for analyzing the influence of saline-alkali stress on the growth of the industrial hemp.
The second embodiment is as follows: in a specific embodiment, the method for evaluating the saline-alkali resistance of industrial hemp in the seedling stage by using a hydroponic method comprises the following steps: 5.75g of ferrous sulfate heptahydrate, 7.45g of disodium ethylene diamine tetraacetate and 1000ml of distilled water, wherein the pH value is 5.5.
The third concrete implementation mode: in a specific embodiment, the method for evaluating the saline-alkali resistance of industrial hemp in the seedling stage by using a hydroponic method comprises the following steps: boric acid 0.77mg/L, manganese sulfate 0.16mg/L, zinc sulfate 0.28mg/L, sodium molybdate 0.025mg/L, copper sulfate 0.062mg/L, cobalt chloride 0.025mg/L, and distilled water 1000mL, with pH 5.5.
The fourth concrete implementation mode: in a first specific embodiment, the method for evaluating the saline-alkali resistance of industrial hemp in the seedling stage by using a hydroponic method comprises the following three steps: the light quantum flux density is 175 +/-2 mu mol.m-2s-1The photoperiod is 14h/10h, the temperature is 21 ℃, the relative humidity is 60%, 100mL of distilled water is supplemented every 2 days, and the nutrient solution is replaced 2 times per week. The limitation of the cultivation conditions is suitable for the growth of industrial hemp. Under the condition, the industrial hemp can grow well, and the phenomena of excessive growth and slow growth caused by uncomfortable conditions are avoided.
The fifth concrete implementation mode: in the fourth step, the agronomic characters are plant height, root length, fresh weight of leaves, fresh weight of roots, fresh weight of stalks, dry weight of leaves, dry weight of stalks and dry weight of roots.
Example 1:
1. seed germination acceleration
Test materials: salt and alkali tolerant industrial hemp variety cannabis sativa No. 1 (provided by Daqing division of the academy of sciences of Heilongjiang province); saline-alkali sensitive type golden knife for industrial hemp variety-15 (provided by Daqing division of the academy of sciences of Heilongjiang province).
Placing 2 varieties of industrial hemp in a culture dish, placing 3 layers of filter paper on the upper part and the lower part, soaking the industrial hemp in distilled water, and placing the industrial hemp in an incubator at the temperature of 25 ℃ for dark culture. Culturing for about 2 days until the bud grows to about 1cm, and transplanting into a culture box.
2. Hydroponic transplanting
The preparation method of the nutrient solution comprises the following steps: 945mg/L of calcium nitrate tetrahydrate, 404mg/L of potassium nitrate, 120mg/L of urea, 230mg/L of ammonium dihydrogen phosphate, 246mg/L of magnesium sulfate, 1ml/L of iron salt solution and 6.5 parts of trace element solution 2ml/L, pH; iron salt solution: 5.75g of ferrous sulfate heptahydrate, 7.45g of disodium ethylene diamine tetraacetate (EDTA.Na) and 1000ml of distilled water, wherein the pH value is 5.5; trace element liquid: boric acid 0.77mg/L, manganese sulfate 0.16mg/L, zinc sulfate 0.28mg/L, sodium molybdate 0.025mg/L, copper sulfate 0.062mg/L, cobalt chloride 0.025mg/L, and distilled water 1000mL, with pH 5.5.
Culturing in 30cm × 20cm × 15cm light-proof plastic box, adding 2/3 times of nutrient solution 1.5L, placing plastic foam with thickness of about 1cm, perforating on the foam, spacing every hole by 3cm, placing small cotton balls and seeds in the holes, and placing in artificial climate box under controllable cultivation condition that light quantum flux density is (175 + -2) μmol · m-2s-1The photoperiod is 14h/10h, the temperature is 21 ℃, and the relative humidity is 60%. Every 2 days, 100mL distilled water was supplemented, and the nutrient solution was changed 2 times per week.
3. Evaluation test of salt base resistance
And about 20 days later, when the seedlings grow to 3 pairs of true leaves, setting 25 seedlings per box, and dividing the seedlings into a control group and a treatment group. Respectively collecting plant samples of golden knife-15 and hemp I, and measuring the plant height, root length, fresh leaf weight, fresh root weight, fresh stem weight, dry leaf weight, dry stem weight, dry root weight and other indexes of the plants as a control. Adding NaHCO into the culture solution3The concentration was 100mM, and the control group was a normal culture medium. The culture medium was changed every 2 days. Up to day 5, surveyAnd (3) the number of dead seedlings is counted, plant samples of golden knife-15 and Cannabis sativa I are collected, and the plant height, the root length, the fresh weight of leaves, the fresh weight of roots, the fresh weight of stalks, the dry weight of leaves, the dry weight of stalks, the dry weight of roots and other indexes are measured and used for analyzing the influence of saline-alkali stress on the growth of the industrial hemp.
4. Analysis of results
TABLE 1 NaHCO3Change of the treated fructus Cannabis
As can be seen from Table 1, NaHCO3Under stress, the plant height, the root length and the dry matter accumulation of the cannabis sativa L.No. 5 days after treatment are all lower than those of a control group under normal conditions by 36.71%, 89.54% and 36.71%, respectively, which shows that NaHCO is used for solving the problem that the plant height, the root length and the dry matter accumulation of the cannabis sativa L.No. 5 days after treatment are all lower than those3Stress inhibited the growth of Cannabis sativa I. NaHCO 23After 5 days of stress, the root-cap ratio of cannabis one is greater than that of the control group, and the results are consistent with previous findings.
TABLE 2 NaHCO3Change of gold knife-15 after treatment
As can be seen from Table 2, NaHCO3Under stress, the plant height, the root length and the dry matter accumulation of the golden knife-15 after 5 days of treatment are respectively reduced by 18.92 percent, 69.49 percent and 61.04 percent compared with the control group under normal conditions, which shows that NaHCO is used3Stress inhibited the growth of golden knife-15. NaHCO 23After 5 days of stress, the root-cap ratio of golden knife-15 was greater than that of the control group, which is consistent with the previous findings.
FIGS. 1 and 2 are schematic representations of the growth of two cannabis varieties, NaHCO3After 5 days of stress, the death rates of the first herb of Cannabis and the second herb of Tanshiya-15 were investigated, and the death rates of the first herb of Cannabis and the second herb of Tanshiya-15 were 67.5% and 82.9%, respectively, which indicates that the disease rate of Cannabis isThe survival rate of the first cutter is 91.18 percent higher than that of the golden cutter-15.
Practice proves that the method can identify the salt and alkali tolerance of main industrial hemp varieties and germplasm resources within 27 days, and the method is simple, reliable, low in cost and high in speed, can improve the accuracy, the scientificity and the reliability of experimental results, and provides a scientific method for evaluating the salt and alkali tolerance of the industrial hemp varieties.
Claims (5)
1. A method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using a water culture method is characterized by comprising the following steps: the method comprises the following steps:
the method comprises the following steps: respectively placing 3 layers of filter paper on the upper and lower parts of different varieties of industrial hemp, soaking the industrial hemp in distilled water, and culturing the industrial hemp in the dark at the temperature of 20-25 ℃ for 2-3 days;
step two: preparing a nutrient solution for water culture, wherein the nutrient solution comprises the following specific components: 945mg/L of calcium nitrate tetrahydrate, 404mg/L of potassium nitrate, 120mg/L of urea, 230mg/L of ammonium dihydrogen phosphate, 246mg/L of magnesium sulfate, 1ml/L of iron salt solution and 6.5 parts of trace element solution 2ml/L, pH;
step three: diluting the concentration of the nutrient solution in the second step to 2/3 times, culturing the industrial hemp obtained in the first step in a light-tight plastic box, filling the culture solution in the plastic box, putting the plastic box into a floating plate, punching the floating plate, wherein the distance between the holes is 2-5 cm, putting small cotton balls and the germinated seeds obtained in the first step into the holes, and putting the small cotton balls and the germinated seeds into an artificial climate box;
step four: evaluation of salt-base tolerance: when 3 pairs of true leaves grow, dividing the seedlings into a control group and a treatment group; respectively collecting plant samples of different varieties as controls, and determining the agronomic characters; adding NaHCO into the culture solution3Making the concentration of the culture medium reach 100mM, and the control group is normal culture medium; changing the culture solution every 2 days; and (5) surveying the number of dead seedlings as long as the day 5, collecting plant samples of different varieties, and determining the agronomic characters for analyzing the influence of saline-alkali stress on the growth of the industrial hemp.
2. The method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using the hydroponics method as claimed in claim 1, which is characterized in that: in the second step, the formula of the iron salt solution is as follows: 5.75g of ferrous sulfate heptahydrate, 7.45g of disodium ethylene diamine tetraacetate and 1000ml of distilled water, wherein the pH value is 5.5.
3. The method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using the hydroponics method as claimed in claim 1, which is characterized in that: in the second step, the formula of the microelement liquid is as follows: boric acid 0.77mg/L, manganese sulfate 0.16mg/L, zinc sulfate 0.28mg/L, sodium molybdate 0.025mg/L, copper sulfate 0.062mg/L, cobalt chloride 0.025mg/L, and distilled water 1000mL, with pH 5.5.
4. The method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using the hydroponics method as claimed in claim 1, which is characterized in that: in the third step, the cultivation conditions are as follows: the light quantum flux density is 175 +/-2 mu mol.m-2s-1The photoperiod is 14h/10h, the temperature is 21 ℃, the relative humidity is 60%, 100mL of distilled water is supplemented every 2 days, and the nutrient solution is replaced 2 times per week.
5. The method for evaluating the saline-alkali resistance of industrial hemp in seedling stage by using the hydroponics method as claimed in claim 1, which is characterized in that: in the fourth step, the agronomic characters are plant height, root length, fresh weight of leaves, fresh weight of roots, fresh weight of stalks, dry weight of leaves, dry weight of stalks and dry weight of roots.
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