CN110810034A - Cutting seedling method of rhododendron xiangzhou - Google Patents
Cutting seedling method of rhododendron xiangzhou Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention provides a cutting seedling raising method of rhododendron parvifolium, belonging to the technical field of rhododendron planting, comprising seedbed preparation, collection of cutting slips, cutting of the cutting slips, cutting and cutting management, wherein a mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately after the collection and cutting of the cutting slips. The invention can inhibit the xylem of the cutting shoot from being embolized, inhibit the reduction of the hydraulic conductivity, improve the survival rate of the cutting shoot, improve the activity of key respiration enzymes of root system cells of cutting seedling, improve the chlorophyll content in leaves, promote the growth and development of the cutting shoot and improve the quality of seedlings.
Description
Technical Field
The invention belongs to the technical field of rhododendron planting, and particularly relates to a cutting seedling method of rhododendron xiangzhou.
Background
The flowering of the fragrant-cheek rhododendron is pink, the plant is compact, the flower is dense, and the fragrant-cheek rhododendron is suitable for being cultivated in a pot or being cultivated in a field. The young branches are thick and green and loose bristles; the leaves are paper, oval, densely covered with yellow-brown hairs, the length of the leaves is 4.0-5.0cm, the width of the leaves is 1.5-2.0cm, the diameter of the flowers is about 7-10cm, half-petal sleeved flowers are dark pink, the leaves are wide and funnel-shaped, red spots are formed in the corolla split pieces, stamens are partially or completely petaloid, 1 pistil is formed, and stigma is yellow. The propagation mode of the variety is cuttage or grafting propagation, the growth vigor is strong, the planting time is four seasons in a year, the natural flowering phase is planted in open field for 7-11 months, the single flower lasts for 15-20 days, and the variety can be cultivated into the flower for the day of national day and Christmas.
The natural hybridization probability of rhododendron is high, the gene purity is difficult to maintain in seeding and seedling raising, the requirements of tissue culture rapid propagation on facility conditions are strict, the technology is complex, the cost is high, and the effective popularization is difficult; the cutting propagation can not only keep the excellent characters of the female parent, shorten the seedling raising time and advance the flowering phase, but also has low cost and easy popularization. At present, the cutting seedling raising technology is widely applied to the large-scale production of the deciduous leaves, the semievergreen rhododendrons and the varieties thereof. The principle of cuttage is that the young somatic cells of the plant have potential totipotency, each somatic cell contains complete sets of genetic information, and the whole individual can be reconstructed on the basis of artificial induced division and differentiation. Comprises hardwood cutting, twig cutting and root cutting. In the cutting technology, many influencing factors are included, such as cutting medium, cutting time, cutting specification, external environment factors and the like.
In the prior art, for example, the patent of the invention in China with the publication number of CN 105210753B, the invention discloses a rhododendron giganteum cuttage technology, which comprises the following steps: mixing the mature soil, the raw soil and the humus soil according to the ratio of 1:1, and then covering with a film; 3-4 days before cuttage, removing the film from the substrate (after mixing), putting the substrate (after mixing) into a container bag, and placing the container bag on a seedling bed; collecting current-year branches of rhododendron with green color (or green color in white), reserving 10-12cm of top stems, reserving 3-5 leaves, and reserving 1/4 parts of whole leaves of each leaf; before cuttage, thoroughly watering a container bag on a seedling bed with clear water, inserting branches treated with rooting powder into the bag, watering the branches with clear water once, and watering carbendazim; building a small arched shed on the cuttage seedling bed, covering the small arched shed with a film, and covering a sunshade net on the arched shed to keep the temperature and the relative humidity of air in the shed; and opening a small arch shed after 1 month to observe the cuttage condition. The invention improves the success rate of cutting propagation of rhododendron by reasonable selection of culture medium and special treatment of cutting branches.
Disclosure of Invention
The invention aims to provide a cutting seedling raising method of rhododendron parvifolium, which can inhibit the xylem of a cutting shoot from being embolized, inhibit the reduction of the hydraulic conductivity, improve the survival rate of the cutting shoot, improve the activity of key respiration enzymes of root system cells for cutting seedling raising, improve the chlorophyll content in leaves, promote the growth and development of the cutting shoot and improve the quality of seedlings.
The technical scheme adopted by the invention for realizing the purpose is as follows:
the cutting seedling method of the rhododendron parvifolium is characterized by comprising the following steps:
s1, preparing a seedbed: disinfecting the substrate with 0.1-0.3% carbendazim, and laying a seedbed;
s2, collection of cutting shoots: selecting a stock plant which is grown vigorously for 2-4 years and has no diseases and insect pests, pruning the stock plant in 1-3 months, and selecting a current-year semi-lignified twig from the stock plant as a cutting shoot in cloudy days or in the early morning in 5-6 months or 9-10 months;
s3, cutting the cutting slips: pruning the cutting branches, wherein the length of the pruned branches is 7-9cm, 3-5 leaves are reserved at the top ends of the branches, and the size of the leaves is 3-4.5 cm;
s4, cuttage: disinfecting the substrate by 0.1-0.3 wt% of potassium permanganate 18-30h before cuttage, watering the substrate thoroughly by water 4-8h before cuttage, and immediately cuttage after the cutting slips are soaked in hormone solution for 5-8 min;
s5, cuttage management: the temperature of 25-35 days before cuttage is not more than 35 ℃, the relative humidity of air is kept at 80-90%, and the shading rate is kept at 75-85%;
wherein, after the collection of the cutting slips in the step S2, the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately after the cutting slips in the step S3. The transpiration causes the water in the branches and leaves of the plants to be continuously lost to the surrounding environment, and the water column has certain tension or negative pressure due to the transpiration force generated by the cohesive force of the water in the xylem conduit. Therefore, when the cutting is cut off from the parent, the water in the xylem duct can be quickly retracted due to the release of the tension, and the cutting is in a water loss state all the time. If the cutting is not carried out in time or the cutting is carried out in sand after the cutting, a large amount of moisture is lost when the cutting slips are exposed in the air. The cutting has no root system, can not absorb a large amount of water from the soil, can only absorb a small amount of water through the cut and the skin hole, and the loss of the water in the cutting body is not beneficial to the formation of the adventitious root, so the cutting is dead when the water loss is serious. Before the cutting has the water absorption function, the cutting can not root and sprout because of the low water content, so the survival rate is low. After the cutting slips are cut off from the matrix and cut, the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped at once, so that the loss of water of the cutting slips in the interval process before cutting can be reduced, the reduction of the water potential of xylem is inhibited, the occurrence of embolization of the xylem of the cutting slips is inhibited, the reduction of the water conductivity is inhibited, the survival rate of the cutting slips is further improved, and the growth and development of the cutting slips are promoted. And the method is simple and convenient, easy to operate, low in cost and beneficial to large-scale cutting seedling raising of the rhododendron odoratum.
In some embodiments, the substrate in step S1 includes river sand, peat, vermiculite, rock wool, quartz sand, mushroom dregs.
In some embodiments, the cutting method of the cutting shoot in step S3 is selected from one of the following: right angle cutting, oblique cutting, double-sided reverse cutting, and necking.
In some embodiments, the hormone solution in step S4 includes 500-800mg/L IBA.
In some embodiments, the matrix further comprises poplar flower extract. The poplar flower extract is added into the substrate, so that the activity of the key respiration enzyme of root system cells of the cutting seedling can be improved, the cell respiration is promoted, the energy metabolism is improved, the absorption of mineral elements is promoted, the chlorophyll synthesis is promoted, the chlorophyll content in leaves is improved, the photosynthesis is enhanced, the growth and development of the root, stem and leaf morphological indexes of the cutting seedling are promoted, and the seedling quality is improved.
In some embodiments, the poplar flower extract is present in an amount of 1-3 wt%.
The invention provides a cutting medium which is characterized by comprising river sand, peat, vermiculite, rock wool, quartz sand, mushroom dregs and poplar flower extracts.
The invention provides application of sodium alginate and sodium propyl p-hydroxybenzoate in improving the water conductivity of a rhododendron gill cutting slip.
The invention provides application of a cutting seedling method in industrialized seedling of rhododendron parvifolium.
The invention also provides application of the cutting medium in improving the quality of the rhododendron parvifolium cutting seedlings.
The invention has the beneficial effects that:
1) according to the method, after the cutting slips are cut off from the matrix and cut, the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately, so that the water loss of the cutting slips in the interval process before cutting can be reduced, the reduction of the water potential of xylem is inhibited, the occurrence of embolization of xylem of the cutting slips is inhibited, the reduction of the water conductivity is inhibited, the survival rate of the cutting slips is further improved, and the growth and development of the cutting slips are promoted;
2) according to the invention, the poplar flower extract is added into the substrate, so that the activity of the key respiration enzyme of root system cells of the cutting seedling can be improved, the cell respiration is promoted, the energy metabolism is improved, the absorption of mineral elements is promoted, the synthesis of chlorophyll is promoted, the chlorophyll content in leaves is improved, the photosynthesis is enhanced, the growth and development of root, stem and leaf morphological indexes of the cutting seedling are promoted, and the seedling quality is improved;
3) the cutting seedling method provided by the invention has the advantages of simplicity, convenience, easy operation, low cost and high survival rate, and is beneficial to large-scale cutting seedling of the rhododendron parvifolium.
Drawings
FIG. 1 shows the water content and water potential of cuttings in test example 1 of the present invention;
FIG. 2 is the hydraulic conductivity of the cutting in test example 1 of the present invention;
FIG. 3 shows HK, PEK, PK, ICDH activities of the root line cells of the cutting seedlings in test example 2 of the present invention;
FIG. 4 shows the contents of N, Mg and Fe in the cutting seedlings in test example 3 of the present invention;
FIG. 5 shows chlorophyll a, chlorophyll b and total chlorophyll contents of the cutting seedlings in test example 3 of the present invention;
FIG. 6 shows the plant height, the number of leaves, the number of roots and the survival rate of the seedlings in test example 4 of the present invention;
FIG. 7 shows the stem thickness, leaf area, average root length and total biomass of the seedling in test example 4 of the present invention.
Detailed Description
Unless otherwise indicated, all publications, patent applications, patents, and other references mentioned herein are incorporated by reference in their entirety as if set forth in their entirety.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. In case of conflict, the present specification, including definitions, will control.
When an amount, concentration, or other value or parameter is given as either a range, preferred range, or a list of upper preferable values and lower preferable values, this is to be understood as specifically disclosing all ranges formed from any pair of any larger range limit or preferred value and any smaller range limit or preferred value, regardless of whether ranges are separately disclosed. For example, when a range of "1 to 5" is described, the described range should be construed as including ranges of "1 to 4", "1 to 3", "1 to 2 and 4 to 5", "1 to 3 and 5", and the like. Where numerical ranges are described herein, unless otherwise stated, the stated ranges are intended to include the endpoints of the ranges and all integers and fractions within the ranges.
In addition, the words "a" and "an" preceding an element or component of the invention are intended to mean no limitation on the number of times that the element or component appears (i.e., occurs). Thus, "a" or "an" should be understood to include one or at least one and the singular forms of an element or component also include the plural unless the singular is explicitly stated.
Embodiments of the present invention, including embodiments of the invention described in the summary section and any other embodiments described herein below, can be combined arbitrarily.
The present invention is described in detail below.
The cutting seedling method of the rhododendron parvifolium is characterized by comprising the following steps:
s1, preparing a seedbed: after sterilizing the substrate with 0.1-0.3%, (preferably, e.g., 0.1%, 0.12%, 0.15%, 0.24%, 0.28%, 0.3%, etc.) carbendazim, the seedbed is laid;
s2, collection of cutting shoots: selecting a stock plant which grows vigorously and has no plant diseases and insect pests for 2-4 years (preferably, for example, 2 years, 2.5 years, 3 years, 4 years and the like), pruning the stock plant in 1-3 months (preferably, for example, in the late 1 month, in the early 2 months, in the early 3 months and the like), and selecting a current-year semi-lignified tender branch from the stock plant in the shade or early morning as a cutting shoot in 5-6 months (preferably, for example, in the early 5 months, in the middle 6 months, in the late 6 months and the like) or in 9-10 months (preferably, for example, in the middle 9 months, in the late 10 months and the like);
s3, cutting the cutting slips: pruning shoot of the cutting shoot, wherein the length of the pruned shoot is 7-9cm (preferably, for example, 7cm, 7.6cm, 7.9cm, 8cm, 8.8cm, 9cm and the like), the top end of the shoot retains 3-5 (preferably, for example, 3, 4 and 5) leaves, and the size of the leaf is 3-4.5cm (preferably, for example, 3cm, 3.6cm, 4.2cm, 4.5cm and the like);
s4, cuttage: disinfecting the substrate with 0.1-0.3 wt% (preferably, e.g., 0.1 wt%, 0.18 wt%, 0.2 wt%, 0.25 wt%, 0.3 wt% etc.) of potassium permanganate 18-30h (preferably, e.g., 18h, 22h, 24h, 25h, 28h, 30h, etc.) before cutting, watering the substrate with water 4-8h (preferably, e.g., 4h, 5h, 6h, 7h, 8h, etc.) before cutting, and cutting immediately after soaking the cutting spike in a hormone solution for 5-8min (preferably, e.g., 5min, 6min, 7min, 8min, etc.);
s5, cuttage management: after cutting, the temperature of the first 25-35d (preferably, for example, 25d, 28d, 30d, 32d, 35d and the like) is not more than 35 ℃ (preferably, for example, 28 ℃, 30 ℃, 32 ℃, 35 ℃ and the like), the relative air humidity is kept at 80-90% (preferably, for example, 80%, 82%, 85%, 90% and the like), the light shading rate is kept at 75-85% (preferably, for example, 75%, 76%, 78%, 80%, 85% and the like), and weeding is performed in time;
wherein, after the collection of the cutting slips in the step S2, the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately after the cutting slips in the step S3. Preferably, the ratio of sodium alginate: sodium propyl p-hydroxybenzoate: the mass ratio of water is 5-6:1-2:300-450, more preferably, the mass ratio of sodium alginate: sodium propyl p-hydroxybenzoate: the mass ratio of water is 5:1: 350. The transpiration causes the water in the branches and leaves of the plants to be continuously lost to the surrounding environment, and the water column has certain tension or negative pressure due to the transpiration force generated by the cohesive force of the water in the xylem conduit. Therefore, when the cutting is cut off from the parent, the water in the xylem duct can be quickly retracted due to the release of the tension, and the cutting is in a water loss state all the time. If the cutting is not carried out in time or the cutting is carried out in sand after the cutting, a large amount of moisture is lost when the cutting slips are exposed in the air. The cutting has no root system, can not absorb a large amount of water from the soil, can only absorb a small amount of water through the cut and the skin hole, and the loss of the water in the cutting body is not beneficial to the formation of the adventitious root, so the cutting is dead when the water loss is serious. Before the cutting has the water absorption function, the cutting can not root and sprout because of the low water content, so the survival rate is low. After the cutting slips are cut off from the matrix and cut, the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped at once, so that the loss of water of the cutting slips in the interval process before cutting can be reduced, the reduction of the water potential of xylem is inhibited, the occurrence of embolization of the xylem of the cutting slips is inhibited, the reduction of the water conductivity is inhibited, the survival rate of the cutting slips is further improved, and the growth and development of the cutting slips are promoted. And the method is simple and convenient, easy to operate, low in cost and beneficial to large-scale cutting seedling raising of the rhododendron odoratum.
In some embodiments, the substrate in step S1 includes river sand, peat, vermiculite, rock wool, quartz sand, mushroom dregs. Preferably, the mass ratio of the river sand, the peat, the vermiculite, the rock wool, the quartz sand, the mushroom dregs and the poplar flower extract is 7-10:12-18:10-13:11-13:8-11:18-25, and more preferably, the mass ratio of the river sand, the peat, the vermiculite, the rock wool, the quartz sand and the mushroom dregs is 8:12:11:12:10: 22.
In some embodiments, the cutting method of the cutting shoot in step S3 is selected from one of the following: right angle cutting, oblique cutting, double-sided reverse cutting, and necking. In some embodiments, the cutting method of the cutting slips in the step S3 is a nering type cutting.
In some embodiments, the hormone solution in step S4 includes 500-800mg/L (preferably, e.g., 500mg/L, 680mg/L, 750mg/L, 800mg/L, etc.) IBA.
In some embodiments, the matrix further comprises poplar flower extract. The poplar flower extract is added into the substrate, so that the activity of the key respiration enzyme of root system cells of the cutting seedling can be improved, the cell respiration is promoted, the energy metabolism is improved, the absorption of mineral elements is promoted, the chlorophyll synthesis is promoted, the chlorophyll content in leaves is improved, the photosynthesis is enhanced, the growth and development of the root, stem and leaf morphological indexes of the cutting seedling are promoted, and the seedling quality is improved.
In some embodiments, the poplar flower extract is present in an amount of 1-3 wt%, preferably, e.g., 1 wt%, 1.2 wt%, 1.8 wt%, 2.6 wt%, 3 wt%, etc.
The invention provides a cutting medium which is characterized by comprising river sand, peat, vermiculite, rock wool, quartz sand, mushroom dregs and poplar flower extracts.
The invention provides application of sodium alginate and sodium propyl p-hydroxybenzoate in improving the water conductivity of a rhododendron gill cutting slip.
The invention provides application of a cutting seedling method in industrialized seedling of rhododendron parvifolium.
The invention also provides application of the cutting medium in improving the quality of the rhododendron parvifolium cutting seedlings.
The present invention is further described in detail with reference to the following examples:
example 1:
a cutting seedling method of rhododendron parvifolium comprises the following steps:
preparing a cutting anti-embolism mixture: dissolving 50g of sodium alginate and 10g of sodium propyl p-hydroxybenzoate in 3.5kg of water, stirring to dissolve completely, radiating with 400W microwave for 6min, and performing ultrasonic treatment for 30 min.
Preparing a seedbed: weighing 11kg of river sand, 16kg of peat, 15kg of vermiculite, 16kg of rock wool, 14kg of quartz sand, 27kg of mushroom residue and 1kg of poplar flower extract, stirring uniformly, loading into nutrient bags of 8cm × 11cm in batches, sterilizing with 0.1% of carbendazim, fully mixing uniformly, loading into 32-hole trays (50 × 27 × 10cm), and putting the hole trays in a greenhouse of 50m length and 7m width in order.
Collecting cutting slips: selecting a 3-year-old vigorous and pest-free gill rhododendron stock plant, pruning the stock plant in 1 month, taking clean and sharp branch scissors 8 o' clock before the early morning in 6 months, taking the current-year semi-lignified twig as a cutting shoot from the stock plant, wherein the length of the cutting shoot is about 11cm, and immediately dipping the cutting shoot embolism prevention mixture.
Cutting the cutting slips: pruning the cutting branches, keeping the length of the branches to be about 8cm after pruning, keeping 3 leaves at the top ends of the branches, keeping the size of the leaves to be 3.6-4.5cm, and immediately dipping the cutting anti-embolism mixture.
Cuttage: and (3) disinfecting the substrate by using 0.3 wt% of potassium permanganate 24 hours before cuttage, watering the substrate thoroughly by using water 6 hours before cuttage, soaking the cutting slips in a solution containing 500mg/L IBA for 6min, and then immediately cuttage, wherein the plant spacing of the cuttage is 2cm, the row spacing is 5cm, the depth is 2cm, the front faces of the leaves face uniformly, and the axillary bud direction is vertically upward.
Cuttage management: after cuttage, bamboo strips are used on the bed surface in time to make a small arched shed about 50cm high, an agricultural film is used for sealing covering (the agricultural film is tightened), the periphery is compacted and pressed by bricks or soil, the temperature of the front 30d is not more than 32 ℃, the relative humidity of air is kept at 85%, and the shading rate is kept at 80%. And (4) checking the sealing condition of the agricultural film, repairing the damaged agricultural film in time and replenishing water, and completely and uniformly watering the agricultural film when replenishing water. After 45d of cuttage, one end of the agricultural film can be opened for ventilation.
Comparative example 1:
the cutting anti-embolism mixture does not contain sodium propyl p-hydroxybenzoate, and the rest part is completely consistent with the example 1.
Comparative example 2:
the anti-embolism mixture of the cutting does not contain sodium alginate, and the rest is completely consistent with the example 1.
Comparative example 3:
after the cutting slips were collected and cut, the mixture for preventing the cutting slips from being clogged was not dipped, and the remaining part was completely the same as that of example 1.
Comparative example 4:
the cutting medium does not include poplar flower extract, and the rest is completely the same as example 1.
Comparative example 5:
after the cutting slips were collected and cut, the mixture for preventing the cutting slips from being clogged was not dipped, the cutting medium did not include the poplar flower extract, and the rest was completely the same as in example 1.
Test example 1:
and (3) measuring the water content, water potential and water conductivity of the cutting:
numbering the cutting slips before cutting, measuring the outer diameters of the cutting slips by using a vernier caliper, and measuring the water content of the cutting slips, wherein the measuring step comprises the steps of firstly recording the fresh weight of the cutting slips with the length of 5cm corresponding to each cutting slip, then putting the cutting slips into a blast drier for drying treatment to reach a constant weight, recording the final mass, and calculating the water content, wherein the water content is (the weight after drying-the fresh weight of the cutting slips)/the fresh weight of the cutting slips; the water potential of the cuttings was measured using a model ZLZ-5 pressure chamber produced by Lanzhou university; the hydraulic conductivity of the cuttings was measured by the "washing method". The water content and water potential of the cuttings are shown in FIG. 1. The results of the measurement of the hydraulic conductivity of the cuttings are shown in fig. 2.
As can be seen from fig. 1 and fig. 2, the water content, water potential and water conductivity of the cutting slips in the example 1 and the comparative example 4 are obviously higher than those in the comparative examples 1, 2, 3 and 5, which shows that the mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately after the cutting slips are collected and cut, so that the water loss of the cutting slips in the interval process before cuttage can be reduced, the water potential of xylem is inhibited from being reduced, the xylem of the cutting slips is inhibited from being embolized, and the water conductivity is inhibited from being reduced.
Test example 2:
and (3) measuring the activity of the respiration key enzyme of the root system cells of the cutting seedlings:
1. the activity of Hexokinase (HK) is measured by taking 0.5g of rhododendron parvifolium cutting seedling root system, grinding in 3ml of phosphate buffer (containing 1.4mM β -mercaptoethanol, 0.1g of PVP40,000, pH8.0) in ice bath, centrifuging homogenate at 4 ℃ at 11000rpm for 10min, and measuring the enzyme activity by using a hexokinase detection kit.
2. Determination of Phosphofructokinase (PEK) activity: 1.0g of the root of the Rhododendron parvifolium cutting seedling was ground in 5ml of LTris-HCl (pH7.50.1mol/L) in an ice bath, and the homogenate was centrifuged at 4000rpm at 4 ℃ for 30 min. The enzyme activity is determined by using a plant phosphofructokinase detection kit.
3. Determination of Pyruvate Kinase (PK) activity: taking 3g of rhododendron parvifolium cutting seedling root in 10mL of Tris-HCl buffer solution (containing 10% glycerol, 15mM mercaptoethanol, 1mM EDTA, 5mM MgCl)2) Grinding in ice bath. The homogenate was filtered through 4 layers of gauze and centrifuged at 11000rpm for 15 min. The enzyme activity was measured with pyruvate kinase assay kit.
4. The determination of isocitrate dehydrogenase activity (ICDH) is carried out by taking 0.5g of gill rhododendron cutting seedling root system, adding 3ml of 0.03mol/LpH7.5 potassium phosphate (containing 1.1M leupeptin, 2mM β -mercaptoethanol and 1% PVP) buffer solution, ice-dissolving and grinding, centrifuging at 11000rpm for 15min, taking supernatant for enzyme activity determination, determining enzyme activity by using isocitrate dehydrogenase detection kit, and determining HK, PEK, PK and ICDH activity of cutting seedling root system cell with result shown in figure 3.
As can be seen from fig. 3, the HK, PEK, PK, and ICDH activities of the root cells of the cutting seedlings in example 1, comparative example 2, and comparative example 3 are significantly higher than those in comparative example 4 and comparative example 5, which shows that the addition of the poplar flower extract to the cutting medium can improve the activity of the respiration key enzyme of the root cells of the cutting seedlings and promote the respiration of the cells.
Test example 3:
1. and (3) measuring the mineral element content of the cutting seedlings:
the test mainly measures the contents of key mineral elements N, Mg and Fe which influence chlorophyll synthesis. Drying root systems of the cutting seedlings of the rhododendron parvifolium, grinding, and sieving to be tested.
1) And (3) determination of N content: 0.25g of the sample was weighed into a 100mL glass digestion tube, and 5mL of concentrated sulfuric acid (H) was added2SO4AR, 98%) was shaken on a vortex shaker and allowed to stand overnight for use, while 2 blanks (no sample) were made for each batch. The digestion furnace is cooledThe temperature is set to 375 ℃, after the temperature is reached, the sample is heated and boiled for 30min, then taken down for cooling for 30min (when the sample is cooled to 15min, 5mL of hydrogen peroxide is added, and the sample is shaken and evenly vibrated), then the sample is put into a boiling furnace for heating and boiling for 30min, and then the sample is taken down for cooling for 30min (when the sample is cooled to 15min, 5mL of hydrogen peroxide is added, and the sample is shaken and evenly vibrated). Repeatedly stewing until the digestion liquid becomes clear and transparent. After the digestion tube is cooled, deionized water with the conductivity of less than 1 mu s/cm is added to the digestion tube, the volume is fixed to 100ml, and the mixture is measured by a continuous flow analyzer after being shaken up.
2) Determination of Mg and Fe content: weighing 0.1g of sample at the bottom of a polyethylene digestion tube to avoid the sample from adhering to the inner wall, and adding 5mL of concentrated nitric acid (HNO)3AR, 65%), then digesting by using a high-flux microwave digestion instrument, transferring the digestion solution to a 25mL volumetric flask after digestion, fixing the volume to 25mL by using deionized water with the conductivity less than 1 mu s/cm, and shaking up to be tested. The Mg content was determined by atomic absorption spectrophotometer. The Fe content was determined by inductively coupled plasma mass spectrometry. The measurement results of the contents of N, Mg and Fe of the cutting seedlings are shown in figure 4.
As can be seen from fig. 4, the contents of N, Mg and Fe of the cutting seedlings in example 1, comparative example 2 and comparative example 3 are higher than those in comparative example 4 and comparative example 5, which shows that the mineral element absorption of the cutting seedlings can be promoted by adding the poplar flower extract to the cutting substrate.
2. Measuring the chlorophyll content of the cutting seedling: cleaning and wiping the leaves, shearing the leaves into thin filaments of about 0.1cm by using scissors, uniformly mixing the thin filaments, avoiding cutting the main veins in the period, weighing 0.1g of the leaves, putting the leaves into a 10mL centrifuge tube, adding 8mL of 80% acetone, uniformly shaking, standing the leaves at room temperature in a dark place, shaking the leaves for 3 to 4 times in the dark place, measuring the leaves by using a UV-2250 spectrophotometer after complete leaching, wherein the measuring wavelengths are 663nm, 645nm and 470nm, and adjusting the leaves to zero by using 80% acetone as a control. And (4) respectively calculating the contents of chlorophyll a, chlorophyll b and total chlorophyll. The measurement results of chlorophyll a, chlorophyll b and total chlorophyll content of the cutting seedlings are shown in figure 5.
As can be seen from fig. 5, the chlorophyll a, chlorophyll b and total chlorophyll contents of the cutting seedlings in example 1, comparative example 2 and comparative example 3 are higher than those in comparative example 4 and comparative example 5, which shows that the addition of the poplar flower extract to the cutting matrix can increase the chlorophyll content in the leaves and enhance photosynthesis.
Test example 4:
measuring seedling growth parameters:
and (4) counting the survival rate after 4 months of cuttage, measuring the plant height, stem thickness, leaf number, leaf area, root number, average root length and total biomass of the nursery stock, and then calculating the average value. Counting the rooting number of the cutting seedlings by adopting a destructive sampling mode, measuring the root length, and calculating the average root length; drying the whole rhododendron simsii plant at 60 ℃ to constant weight. After cooling, the total biomass was determined on an analytical balance. The measurement results of the plant height, the leaf number, the root number and the survival rate of the nursery stock are shown in figure 6. The measurement results of the stem thickness, leaf area, average root length and total biomass of the seedlings are shown in FIG. 7.
As can be seen from fig. 6 and 7, the plant height, the leaf number, the root number, the survival rate, the stem thickness, the leaf area, the average root length and the total biomass of example 1 are obviously higher than those of comparative example 1, comparative example 2, comparative example 3, comparative example 4 and comparative example 5, and the total biomass of comparative example 3 and comparative example 4 are obviously higher than those of comparative example 5, which shows that the mixed solution of sodium alginate and propyl p-hydroxybenzoate is dipped immediately after the cutting shoot is collected and cut, so that the survival rate of the cutting shoot can be improved, and the growth and development of the cutting shoot are promoted; the poplar flower extract is added into the cutting medium, so that the growth and development of root, stem and leaf morphological indexes of the cutting seedlings can be promoted, and the quality of the seedlings is improved.
Conventional techniques in the above embodiments are known to those skilled in the art, and therefore, will not be described in detail herein.
The above embodiments are merely illustrative, and not restrictive, and those skilled in the art can make various changes and modifications without departing from the spirit and scope of the invention. Therefore, all equivalent technical solutions also belong to the scope of the present invention, and the protection scope of the present invention should be defined by the claims.
Claims (10)
1. A cutting seedling method of rhododendron parvifolium is characterized by comprising the following steps:
s1, preparing a seedbed: disinfecting the substrate with 0.1-0.3% carbendazim, and laying a seedbed;
s2, collection of cutting shoots: selecting a stock plant which is grown vigorously for 2-4 years and has no diseases and insect pests, pruning the stock plant in 1-3 months, and selecting a current-year semi-lignified twig from the stock plant as a cutting shoot in cloudy days or in the early morning in 5-6 months or 9-10 months;
s3, cutting the cutting slips: pruning the cutting branches, wherein the length of the pruned branches is 7-9cm, 3-5 leaves are reserved at the top ends of the branches, and the size of the leaves is 3-4.5 cm;
s4, cuttage: disinfecting the substrate by 0.1-0.3 wt% of potassium permanganate 18-30h before cuttage, watering the substrate thoroughly by water 4-8h before cuttage, and immediately cuttage after the cutting slips are soaked in hormone solution for 5-8 min;
s5, cuttage management: the temperature of 25-35 days before cuttage is not more than 35 ℃, the relative humidity of air is kept at 80-90%, and the shading rate is kept at 75-85%;
and after the cutting slips are collected in the step S2, a mixed solution of sodium alginate and sodium propyl p-hydroxybenzoate is dipped immediately after the cutting slips are cut in the step S3.
2. The cutting seedling raising method according to claim 1, characterized in that: the substrate in the step S1 comprises river sand, peat, vermiculite, rock wool, quartz sand and mushroom residue.
3. The cutting seedling raising method according to claim 1, characterized in that: the cutting method of the cutting slips in the step S3 is selected from one of the following methods: right angle cutting, oblique cutting, double-sided reverse cutting, and necking.
4. The cutting seedling raising method according to claim 1, characterized in that: the hormone solution in the step S4 comprises 500-800mg/L IBA. .
5. The cutting seedling raising method according to claim 2, characterized in that: the matrix also comprises poplar flower extract.
6. The cutting seedling raising method according to claim 5, characterized in that: the content of the poplar flower extract is 1-3 wt%.
7. A cutting medium is characterized by comprising river sand, peat, vermiculite, rock wool, quartz sand, mushroom dregs and poplar flower extracts.
8. The application of sodium alginate and sodium propyl p-hydroxybenzoate in improving the water conductivity of the rhododendron gill cutting slips.
9. Use of the cutting seedling method of any one of claims 1 to 6 in industrialized seedling of rhododendron parvifolium.
10. Use of the cutting substrate of claim 7 for improving the quality of rhododendron parvifolium cutting seedlings.
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