CN114793530A - A kind of method that utilizes seaweed extract to promote cassia seed germination and growth - Google Patents

Abstract

The invention discloses a method for promoting germination and growth of cassia seed seeds by using seaweed extract. Belongs to the technical field of biology, and comprises the following specific operation steps: step (1), preparing raw materials; step (2), preparing an enteromorpha extracting solution; step (3), performing seed germination experiments; step (4), carrying out a cassia seed seedling growth experiment; and (5) carrying out data statistics and analysis. The seaweed extract is a safe and potential multifunctional agricultural biofertilizer; large algal blooms such as green tide, golden tide and the like burst to provide sufficient raw materials for preparing the seaweed extract; the invention provides a new way for further development and utilization of green tide outbreak enteromorpha; the method for preparing the seaweed extract has low cost, and can prepare a large amount of seaweed extract in a short time; compared with the use of chemical fertilizers and pesticides, the seaweed extract is green and has no residue.

Description

A kind of method that utilizes seaweed extract to promote cassia seed germination and growth

technical field

The invention belongs to the field of biotechnology, and relates to a method for promoting the germination of cassia seeds; in particular, to a method for promoting the germination and growth of cassia seeds by utilizing seaweed extract.

Background technique

As a commonly used traditional Chinese medicine, cassia seed is sweet, salty, bitter, slightly cold; it has the functions of clearing heat and improving eyesight, moistening the intestines and laxative, etc. It is used for red eyes and astringent pain, shyness and tears, headache, dizziness, dark eyes, constipation; modern Medical research has shown that cassia seeds have the functions of lowering blood pressure, regulating lipids, protecting liver and improving kidney function. With in-depth research on ingredients, pharmacological effects and clinical effects, cassia seeds have been widely used in the fields of medicine and food. Methods for promoting seed germination include using plant growth regulators such as gibberellin (GA3), indoleacetic acid (IAA), naphthaleneacetic acid (NAA), 6-benzyl adenine (6-BA), kinetin (KT), etc.; Environmental factors such as temperature and salinity promote seed germination.

Macro seaweed is rich in various compounds such as proteins, lipids, phytohormones, carbohydrates, amino acids, osmoprotectants, antibacterial compounds and minerals; the demand for seaweed extracts in agriculture has been greatly stimulated by the demand for organic farming and organic food Such as promoting seed germination, enhancing plant health and growth, improving water and nutrient absorption, antifreeze and salt tolerance, biological control and resistance to plant pathogens, remediation and fertilization of pollutants in polluted soil.

my country's latest relevant policies have formulated the maximum residue limits of 548 pesticides in food, of which 28 are plant growth regulators, accounting for more than 50% of the registered plant growth regulators in my country. Plant growth regulators are frequently used in the process of crop planting, and the long-term unreasonable use, coupled with the slow degradation of some plant growth regulators and the characteristics of easy residues, will inevitably lead to the residues of plant growth regulators. At present, there have been many reports on the residual status of plant growth regulators in crops, among which the detection rate of paclobutrazol is generally high. As a new type of green multifunctional biological fertilizer, seaweed extract can reduce the use and residues of chemical fertilizers and pesticides, and at the same time provide a new way for the further development and utilization of Ulva and green tide outbreak species.

Seaweed extract can be used as a safe and potential multifunctional agricultural bio-fertilizer, and the use of seaweed extract as bio-fertilizer may help alleviate the adverse effects of major nutrient deficiencies and reduce the use of chemical fertilizers; at the same time, proliferate for green tide outbreaks provide new avenues for further development and utilization.

SUMMARY OF THE INVENTION

Purpose of the invention: The purpose of the present invention is to provide a method for promoting the germination and growth of Cassia seeds by utilizing seaweed extract; It may help to alleviate the adverse effects of major nutrient deficiencies and reduce the use of chemical fertilizers; at the same time, it provides a new way for the further development and utilization of E. prolifera.

Technical scheme: a method for promoting the germination and growth of cassia seeds by utilizing seaweed extracts of the present invention, utilizing seaweed extracts to promote the germination of cassia seeds, and the specific operation steps are as follows:

The preparation of step (1), raw material; With seaweed (Prolifera, Ulva) as donor plant material, with cassia seed as bioassay material, for subsequent use;

Step (2), preparing the seaweed extract, using physical methods to break the collected seaweed to extract the seaweed extract, specifically: rinse the collected seaweed with clean water, then wash with pure water 3 times, and place it at 70 ° C Dry in an oven to constant weight, crush the dried seaweed into powder in a wall breaker, which is seaweed powder; add 100 g of seaweed powder to 1000 mL of 50% ethanol, use an ultrasonic cleaning machine for ultrasonic extraction for 2 hours, and place at 4°C for 24 hours; Centrifuge at 4250 rpm for 15 minutes, take the supernatant and filter it with Whatman No. 1 filter paper; extract the lower powder residue once by the above method, and combine the two extraction filtrates; evaporate the filtrate to 100 mL on a rotary evaporator to obtain 1 g/ml seaweed extract , and then serially diluted to obtain 0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract, refrigerated at 4°C for later use;

Step (3), seed germination experiment, its operation method is:

Select cassia seeds, break dormancy with 4% sodium hypochlorite, and soak in the extracted seaweed extract;

Specifically: select plump and healthy cassia seeds, soak and disinfect them with 4% sodium hypochlorite for 10 minutes, and then wash them with pure water for 3-4 times; add the washed seeds to four concentrations of seaweed extract, and pure water as a control group for soaking 24h, the submerged seeds are subject to 5cm, and then washed with pure water; the treated cassia seeds are placed in a petri dish with two layers of filter paper, and the petri dish is cleaned in advance, sterilized in an autoclave, and dried in an oven; each culture 100 seeds were added to 0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract and pure water, and each treatment was repeated 3 times; after that, an appropriate volume of Algae extract and pure water, filter paper was changed every three days; culture conditions were day and night temperature 25°C/20°C, photoperiod 14h/10h, light intensity 100μmol photons m ^-2 s ^-1 .;

Record the germination of the seeds at a fixed time every day, and the seed germination experiment is regarded as the end of the seed germination experiment when the number of germinated seeds does not increase for 48 consecutive hours; the germination potential, germination index, vigor index, and germination rate are determined; the relevant calculation formulas are as follows:

Germination potential: the percentage of seeds germinated within the initial recording time to the number of seeds tested;

Germination index=∑(G _t /D _t ), where: G _t is the germination number within t time, and D _t is the corresponding germination days;

Vitality index=germination index×plant weight (g);

Germination rate (%) = germination number ÷ inoculation number × 100;

Step (4), cassia seedling growth experiment, the cassia seed soaked by seaweed extract is planted in the seedling tray, watered 2 times every day, after the test starts two weeks, each process selects the consistent cassia seedling of growth to carry out chlorophyll, plant height, Determination of root length, specifically;

The cassia seeds were soaked in seaweed extract for 24 hours and planted in seedling trays, watered twice a day, 10 ml per plant, and 3 replicates were set for each treatment, each replicate was 3 plug trays, arranged randomly; two weeks after the start of the experiment, each For each treatment, 10 cassia seedlings with consistent growth were selected for the determination of various indicators;

Chla (mg/g FW)=13.95* _OD665-6.88 * _OD649 ;

Chlb(mg/g FW)=24.96* _OD649-7.32 * _OD665 ;

Plant height (cm): measure the distance from the open field to the top of the seedling to the root when the seedling is upright;

Main root length (cm): measure the distance from the rhizome to the main root tip with a vernier caliper;

(5), data statistics and analysis; all data were sorted and preliminarily analyzed with excel; one-way ANOVA analysis of variance, LSD multiple comparison and Waller-Duncan test were performed with SPSS, and Origin was used to draw histograms.

Beneficial effects: Compared with the prior art, the present invention has the following features: 1. The seaweed extract is a safe and potential multifunctional agricultural biological fertilizer; The seaweed extract provides sufficient raw materials; 3. Provides a new way for the further development and utilization of the green tide outbreak species Prolifera; 4. The cost of preparing the seaweed extract is low, and a large amount of seaweed extract can be prepared in a short time; 5. Compared with Chemical fertilizers and pesticides are used, and the seaweed extract is green and has no residue.

Description of drawings

Fig. 1 is the preparation flow chart of the present invention;

Fig. 2 is the contrast diagram of the influence of different seaweed extracts on cassia plant leaf Chla in the embodiment of the present invention;

Fig. 3 is the contrast diagram of the influence of different concentrations of seaweed extracts on cassia plant leaf Chlb in the embodiment of the present invention;

Fig. 4 is the contrast diagram of the influence of different concentrations of seaweed extracts on the plant height of Cassia in the embodiment of the present invention;

Figure 5 is a comparison diagram of the effects of different concentrations of seaweed extracts on the root length of cassia seeds in the embodiment of the present invention.

Detailed ways

The present invention will be further described below in conjunction with the accompanying drawings and embodiments.

A method for promoting the germination and growth of cassia seeds by utilizing the seaweed extract of the present invention utilizes the seaweed extract to promote the germination of the cassia seeds, and the specific operation steps are as follows:

The preparation of step (1), raw material; With seaweed (Prolifera, Ulva) as donor plant material, with cassia seed as bioassay material, for subsequent use;

Step (2), preparing the seaweed extract, using physical methods to break the collected seaweed to extract the seaweed extract, specifically: rinse the collected seaweed with clean water, then wash with pure water 3 times, and place it at 70 ° C Dry in an oven to constant weight, crush the dried seaweed into powder in a wall breaker, which is seaweed powder; add 100 g of seaweed powder to 1000 mL of 50% ethanol, use an ultrasonic cleaning machine for ultrasonic extraction for 2 hours, and place at 4°C for 24 hours; Centrifuge at 4250 rpm for 15 minutes, take the supernatant and filter it with Whatman No. 1 filter paper; extract the lower powder residue once by the above method, and combine the two extraction filtrates; evaporate the filtrate to 100 mL on a rotary evaporator to obtain 1 g/ml seaweed extract , and then serially diluted to obtain 0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract, refrigerated at 4°C for later use;

Step (3), seed germination experiment, its operation method is:

Cassia seeds were selected, 4% sodium hypochlorite to break dormancy, and soaked in the extracted seaweed extract.

Specifically: select plump and healthy cassia seeds, soak and disinfect them with 4% sodium hypochlorite for 10 minutes, and then wash them with pure water for 3-4 times; add the washed seeds to four concentrations of seaweed extract, and pure water as a control group for soaking 24h, the submerged seeds are subject to 5cm, and then washed with pure water; the treated cassia seeds are placed in a petri dish with two layers of filter paper, and the petri dish is cleaned in advance, sterilized in an autoclave, and dried in an oven; each culture 100 seeds were added to 0.4g/ml, 0.2g/ml, 0.1g/ml, 0.05g/ml seaweed extract and pure water, and each treatment was repeated 3 times; after that, an appropriate volume of Algae extract and pure water, filter paper was changed every three days; culture conditions were day and night temperature 25°C/20°C, photoperiod 14h/10h, light intensity 100μmol photons m ^-2 s ^-1 .;

Record the germination of the seeds at a fixed time every day, and the seed germination experiment is regarded as the end of the seed germination experiment when the number of germinated seeds does not increase for 48 consecutive hours; the germination potential, germination index, vigor index, and germination rate are determined; the relevant calculation formulas are as follows:

Germination potential: the percentage of seeds germinated within the initial recording time to the number of seeds tested;

Germination index=∑(G _t /D _t ), where: G _t is the germination number within t time, and D _t is the corresponding germination days;

Vitality index=germination index×plant weight (g);

Germination rate (%) = germination number ÷ inoculation number × 100;

Step (4), cassia seedling growth experiment, the cassia seed soaked by seaweed extract is planted in the seedling tray, watered 2 times every day, after the test starts two weeks, each process selects the consistent cassia seedling of growth to carry out chlorophyll, plant height, Determination of root length, specifically;

The cassia seeds were soaked in seaweed extract for 24 hours and planted in seedling trays, watered twice a day, 10 ml per plant, and 3 replicates were set for each treatment, each replicate was 3 plug trays, arranged randomly; two weeks after the start of the experiment, each For each treatment, 10 cassia seedlings with consistent growth were selected for the determination of various indicators;

Chla (mg/g FW)=13.95* _OD665-6.88 * _OD649 ;

Chlb(mg/g FW)=24.96* _OD649-7.32 * _OD665 ;

Plant height (cm): measure the distance from the open field to the top of the seedling to the root when the seedling is upright;

Main root length (cm): measure the distance from the rhizome to the main root tip with a vernier caliper;

(5), data statistics and analysis; all data are sorted and preliminarily analyzed with excel; one-way ANOVA analysis of variance and LSD multiple comparison and Waller-Duncan test are performed with SPSS, and Origin is used to draw histograms; as shown in Table 1 and Table 1 2 is shown,

Table 1 Measurement results of pH and conductivity of seaweed extracts with different concentrations

The pH of the seaweed extract was mostly lower than that of the control group. With the increase of the concentration of the seaweed extract, the conductivity also showed an obvious multiple relationship. The conductivity of the seaweed extract at any concentration was higher than that of the control group.

Table 2 Effects of different concentrations of seaweed extract on germination of cassia seeds

Compared with the control group, except for the 0.4% Ulva extract, which has no significant difference, other different concentrations of seaweed extracts can improve the germination potential of cassia seeds; different concentrations of the cassia extract can mention the cassia seeds compared with the control group. Germination index, 0.05% Ulva extract significantly improved the germination index of cassia seeds, and 0.1%, 0.2%, 0.4% had no significant difference compared with the control group; all in all, the proliferative moss extract in promoting the germination of cassia seeds was better than the Ulva extract Among them, the 0.05% prolifera extract had the best effect on promoting the germination of cassia seeds.

The specific embodiment is shown in Figures 2-3. It can be seen from Figures 2 and 3 that, compared with the clear water control group, the application of seaweed extract to water cassia seedlings can effectively promote the generation of chlorophyll content. Compared with the control group, the chla content increased by 128.10%, 98.53%, 248.12%, 183.49% in 0.05%, 0.1%, 0.1%, 0.2% Ulva extract, respectively. %, the content of chlb increased by 79.72%, 200.83%, 127.87% and 91.67% respectively, showing a significant correlation.

It can be seen from Figure 4 that, compared with the control group, the plant height of the applied seaweed extract is higher than that of the control group, but most of them have no significant difference. Compared with the control group, the plant heights increased by 25.8%, 14.49% and 19.14% respectively, showing a significant difference.

As can be seen from Figure 5, compared with the control group, the application of seaweed extract in the root growth, from the root length of 0.05% prolifera extract has a significant increase, other concentrations of seaweed extract have no significant difference.

Combined with the germination test of cassia seeds and the growth test of seedlings, it can be seen that the extract of prolifera and Ulva has a certain promoting effect on the germination of cassia seeds and the growth of seedlings. Among them, the more suitable concentration of seaweed extract is in the range of 0.05%-0.2 %between.

The innovations of the present invention: 1. In addition to physical crushing, the preparation of seaweed extract only needs 50% ethanol extraction, and the process is simple; 2. The effect of seaweed extract on cassia seeds is combined with germination index and growth index. %-0.2%; 3. Provide a new way for the further development and utilization of Prolifera sp.; 4. Seaweed extract can reduce the use of chemical fertilizers and pesticides as a method to promote seed germination.

The above are only preferred embodiments of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and all technical solutions that belong to the idea of the present invention belong to the protection scope of the present invention. It should be pointed out that for those skilled in the art, some improvements and modifications without departing from the principle of the present invention should be regarded as the protection scope of the present invention.

Claims (5)

1. A method for promoting germination and growth of cassia seeds by utilizing seaweed extract is characterized by comprising the following specific operation steps of:

step (1), preparing raw materials;

step (2), preparing seaweed extract;

step (3), performing seed germination experiments;

and (4) carrying out a cassia seed seedling growth experiment.

2. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,

in step (1), the preparation of the material is specifically:

the seaweed is taken as donor plant material, and the cassia seed is taken as test material for standby.

3. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,

in the step (2), the collected seaweed is physically crushed to extract seaweed extract.

4. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,

in the step (3), the specific operation method of the seed germination experiment is as follows:

selecting semen Cassiae seed, breaking dormancy with 4% sodium hypochlorite, and soaking in the extracted seaweed extractive solution.

5. The method for promoting germination and growth of semen Cassiae seeds with Sargassum extract as claimed in claim 1,

in the step (4), the cassia seed seedling growth experiment specifically comprises:

and (3) planting the cassia seeds soaked in the seaweed extracting solution in a seedling culture tray, irrigating for 2 times every day, and after two weeks of the test, selecting cassia seeds with consistent growth vigor for each treatment to measure chlorophyll, plant height and root length.

Images