CN112889567B - Method for obtaining large-batch high-quality explants by using curvicular mangnolia branch sprouting promoting technology and application of method - Google Patents
Method for obtaining large-batch high-quality explants by using curvicular mangnolia branch sprouting promoting technology and application of method Download PDFInfo
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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Abstract
The invention discloses a method for obtaining a large batch of high-quality explants by using a curvatura bicolor branch sprouting promoting technology and application thereof. The method comprises the steps of matrix treatment, pruning treatment, field planting, branch bending, soil covering, germination management and the like. According to the invention, through experimental researches on the bent branch cultivation substrate, the bent branch front branch pruning method, the bent branch method and the like, a bent branch sprouting promoting mode suitable for the castanopsis hystrix is established, the quantity and the quality of the castanopsis hystrix explants can be obviously improved, the problem of insufficient quantity of high-quality castanopsis hystrix explants can be effectively solved, and a feasible theoretical basis and technical support can be provided for preparing the juvenile explants by asexual propagation.
Description
Technical Field
The invention belongs to the technical field of fine variety breeding of forest trees, and particularly relates to a method for obtaining a large batch of high-quality explants by using a red prickle branch sprouting promoting technology and application thereof.
Background
The mangnolia officinalis (Castanopsis hystrix) is evergreen tree of kawasaki of kawasabi, is an important local broad-leaved high-quality precious material and multipurpose tree species in south China, and is mainly distributed in the places of Guangdong, Guangxi, Hainan, Yunnan and the like in China. The red cone has the characteristics of fast growth, wide application, high benefit and the like, and the main stem of the red cone is straight, hard in material, red, strong in corrosion resistance, free of cracking and deformation and easy to process, and can be used for buildings, shipbuilding, high-grade furniture, wooden floors, military supplies, sports equipment and the like. The redcone is more shade-resistant and has good mixed performance, can be planted in pure forest and can also be mixed for afforestation, and is one of the most ideal associated tree species for mixed afforestation with conifer species such as pine, fir and the like.
The current mangosteen artificial forest construction mainly comes from bearing seed seedlings, but the phenomenon of bearing of the mangosteen mother forest in the year and after years is very obvious, and the mangosteen bud seedlings need to be cultured in a nursery land for 2 years to be planted in a mountain, so that the requirement of large-scale forestation is difficult to meet. In recent years, researchers develop a large amount of studies on cutting and tissue culture asexual propagation of castanopsis hystrix, so that fine breeding can be accelerated, the restriction of time and seasonal factors is avoided, the propagation efficiency is greatly improved, the requirements of a large amount of seedlings are met, and the progress is slight. For this reason, the number and quality of explants are important limiting factors for their slow progress. The vigorous growth period of the young growth of the red awl is generally from 3 to 5 months, and the growth is basically stopped from 5 months, so that high-quality explants can be obtained only from 3 to 5 months of each year, and the tissue culture requirement of the current season is met. The proliferation times of the castanopsis hystrix during tissue culture are low, and the growth potential and the survival rate are reduced if the subculture times are too many. At present, the explant acquisition mode in the non-growth vigorous growth stage mainly comprises cutting stake sprout retention, girdling sprout promotion and the like, wherein: the superior tree needs to be felled when the stump is left in the bud, the destructiveness is irreversible, and the cost is high; the initiation of girdling is difficult after the girdling often appears, and the plants die, causing irreparable loss. Therefore, the method is directly applied to the cone rubra and is irrevocable, so that a way for effectively obtaining a large number of explants is urgently found.
Disclosure of Invention
The invention aims to overcome the defects in the prior art and provides a method for obtaining a large batch of high-quality explants by using a curvy red cone branch sprouting promoting technology.
The invention also aims to provide the application of the method for obtaining the large-batch high-quality explants by the curvatura bicolor sprouting promoting technology.
The above object of the present invention is achieved by the following technical solutions:
a method for obtaining a large batch of high-quality explants by using a curvicular rosette sprouting promoting technology comprises the following steps:
s1, matrix treatment: sterilizing the matrix before field planting, covering with a film, lifting the film before field planting, and watering the matrix thoroughly with water;
s2, pruning: pruning trunk branches of the castanopsis hystrix seedlings to obtain pruned castanopsis hystrix seedlings;
s3, field planting: pouring the pruned castanopsis hystrix seedlings obtained in the step S2 into a hole as castanopsis hystrix explant materials, and covering the castanopsis hystrix seedlings with the matrix obtained in the step S1;
s4, branch bending: selecting a part, which is not higher than 15cm away from the base, of the trunk of the pruned castanopsis hystrix seedling as a branch bending part, bending the trunk branch by 90-120 degrees, and fixing the top branch of the pruned castanopsis hystrix seedling;
s5, covering soil: covering the top ends of the branches with a substrate;
s6, germination management: cutting off lateral sprouting branches until the subcutaneous part begins to sprout and root, and the overground part sprouts into juvenile scion strips; fertilizing, preventing diseases, selectively using plant growth regulator to obtain large-scale high-quality explants.
The substrate in the step S1 is preferably a substrate obtained by mixing loess and peat soil according to the mass ratio of 1-2: 1; the matrix is preferably obtained by mixing loess and peat soil according to the mass ratio of 1: 1.
The sterilization described in step S1 is preferably performed using potassium permanganate; more preferably by using a potassium permanganate solution with a concentration of 0.5% (w/v).
The timing of sterilization described in step S1 is preferably sterilization the day before permanent planting.
The time for covering the film in the step S1 is preferably 0.5-1.5 d; more preferably 1 d.
The watering frequency in the step S1 is preferably 2-3 times.
The preferred castanopsis hystrix seedlings described in step S2 are castanopsis hystrix nutrition bag seedlings.
The trimming process described in step S2 is performed in any one of the following modes a to D, preferably mode a:
A. cutting off the top, and cutting off the lateral branches to leave 2-3 lateral branches close to the top;
B. cutting off the top and reserving all lateral branches;
C. without cutting off the top, cutting off the lateral branches and leaving 2-3 lateral branches close to the top;
D. and (4) cutting off all lateral branches and reserving a main trunk and 2-3 leaves.
The castanopsis hystrix explant material described in step S3 is a robust plant, pest and disease free explant material.
The inclined placement in the step S3 means that the cut stem of the castanopsis hystrix seedling is placed at an included angle of 5-30 degrees with the ground; more preferably, the pruned trunk of the castanopsis hystrix seedling is placed at an included angle of 15 degrees with the ground.
The depth of the cavity described in step S3 is preferably not less than 15 cm; preferably 15-25 cm; more preferably 20 cm.
The size of the holes described in step S3 is preferably 30cm × 30cm × 20 cm.
The substrate coverage depth in the step S3 is 15-25 cm above the base of the trimmed castanopsis hystrix seedling; preferably 20 cm.
The bent branch position in the step S4 is preferably a position 5-10 cm away from the base of the trunk of the pruned castanopsis hystrix seedling; more preferably, the trunk of the pruned castanopsis hystrix seedling is 5cm away from the base.
The top branch in the step S4 is preferably processed by any one of the modes 1) to 3); preferred mode 1):
1) the pressure of the branches near the top end is reduced to be not higher than the branch bending part;
2) the branches near the top end are in a platform type;
3) the pressure of the branches near the top end is lower but higher than the bending parts of the branches.
The fixing described in step S4 is preferably with iron rings or wooden stakes.
The matrix in the step S5 is obtained by mixing yellow core soil, peat soil and pond sludge according to the mass ratio of 1-2: 1: 1; preferably, the matrix is obtained by mixing yellow core soil, peat soil and pond sludge according to the mass ratio of 1:1: 1.
The frequency of pruning the lateral shoots from sprouts as described in step S6 is preferably 1 time per month.
The subcutaneous sprouting and rooting time in the step S6 is about 20 days after the lateral sprouting branches are cut off.
The specific steps of fertilization described in step S6 are preferably as follows: and (3) within 1-3 months after subcutaneous sprouting, applying the urea liquid fertilizer for 2-3 times per month, and applying P, K fertilizers while applying the N fertilizer.
The specific operation for preventing diseases described in step S6 is preferably as follows: spraying 1% equivalent Bordeaux mixture or 50% carbendazim solution once a month to prevent diseases.
The plant growth regulator described in step S6 is preferably paclobutrazol.
The use condition of the paclobutrazol is preferably that the paclobutrazol solution with the concentration of 100-150mg/L is applied once after 3 months of subcutaneous sprouting.
The method for obtaining a large number of high-quality explants by the curved branch sprouting promotion technology of the castanopsis hystrix can be widely applied to preparation of clonal seedlings of the castanopsis hystrix, so that a large number of high-quality seedlings are provided for construction of the artificial mangosteen forest.
Compared with the prior art, the invention has the following advantages and beneficial effects:
(1) the invention can solve the bottleneck of asexual propagation difficulty of the castanopsis hystrix, not only can provide high-quality explants with high rejuvenization, but also has high success rate of in vitro culture.
(2) The invention adopts the pruning method of cutting off the top and reserving all/part of the lateral branches, and the sprouting effect is good.
(3) The method selects the position of the trunk of the castanopsis hystrix explant material, which is 5-10 cm away from the base part, as a branch bending part, and the inventor of the application finds that the part is the youngest part of the trunk stage development of the castanopsis hystrix explant material, has strong physiological function, and can firstly obtain nutrients and water delivered from the root, so that the young shoots are drawn and strong, the leaves are large, and the growth is rapid.
(4) According to the method, the trunk branches are bent by 90-120 degrees, the pressure of the branches near the top end is low and is not higher than that of the bent parts of the branches, the germination promoting effect is good, and the number of sprouts is large and robust.
(5) According to the invention, through experimental researches on the bent branch cultivation substrate, the bent branch front branch pruning method, the bent branch method and the like, a bent branch sprouting promoting mode suitable for the castanopsis hystrix is established, the quantity and the quality of the castanopsis hystrix explants can be obviously improved, the problem of insufficient quantity of high-quality castanopsis hystrix explants can be effectively solved, and a feasible theoretical basis and technical support can be provided for preparing the juvenile explants by asexual propagation.
Drawings
FIG. 1 is a schematic representation of the curvy sprout promotion test according to the method described in example 1.
FIG. 2 is a graph showing the effect of bend sprouting by the method described in example 1.
FIG. 3 is a graph showing the effect of different pruning methods on germination effect in example 3.
FIG. 4 is a graph showing the effect of different ways of bending branches on the sprouting effect in example 4.
Detailed Description
The present invention will be described in further detail with reference to examples and drawings, but the present invention is not limited thereto.
The reagents and methods used in the examples are those commonly used in the art, unless otherwise specified, and any insubstantial changes and substitutions made by those skilled in the art based on the present invention are intended to be within the scope of the invention as claimed.
The source of the mangosteen is a mangosteen fine variety base in a longan cave forest farm in Guangdong province; the loess, pond sludge and peat soil are purchased from Guangzhou Fangcun, and the peat soil is 0-10mm in particle size.
Example 1
A method for obtaining a large batch of high-quality explants by using a curvicular rosette sprouting promoting technology is shown in figure 1 and comprises the following steps:
s1, matrix treatment: the matrix is respectively yellow core soil, peat soil and mixed soil (the mixed soil is obtained by mixing the yellow core soil and the peat soil according to the mass ratio of 1: 1), the mixed soil is disinfected by a potassium permanganate solution with the concentration of 0.5% (w/v) one day before field planting, a film is covered for one day, the film is lifted before field planting, water is used for 2-3 times, and the matrix is thoroughly poured;
s2, pruning: cutting off tops of trunk branches of the castanopsis hystrix nutrition bag seedlings, cutting off partial side branches on the trunk branches of the castanopsis hystrix branches, and reserving 2-3 near-top side branches;
s3, field planting: taking the branches with 2-3 near-top side branches obtained by pruning in the step S2 as a castanopsis hystrix explant material (strong plants without diseases and insect pests), and pouring the castanopsis hystrix explant material into a hole (30cm multiplied by 20cm), wherein the included angle between the plants and the ground is 15 degrees, the hole is covered by the matrix in the step S1, and the covering depth is 20cm above the base of the explant; in order to find the most suitable matrix, each matrix is provided with 5 groups of repeats, each group contains 4 strains, and the influence of different matrices on the sprouting number is compared;
s4, branch bending: selecting a part of the trunk of the castanopsis hystrix explant material, which is 5-10 cm away from the base (as close to 5cm as possible during operation), as a branch bending part, bending the trunk branch for 90-120 degrees, simultaneously pressing the top branch of the castanopsis hystrix explant material to be lower than the branch bending part, and fixing the top branch by using an iron ring;
s5, covering soil: covering the top end with a substrate, wherein the substrate is obtained by mixing yellow core soil, peat soil and pond sludge according to the mass ratio of 1:1: 1;
s6, germination management: cutting off lateral shoots of sprouts on a trunk, wherein the sprouts start to sprout and root under the skin and sprout immature scions on the overground part after 20 days 1 time per month; in 1-3 months after subcutaneous sprouting, applying urea liquid fertilizer 2-3 times per month, and applying P, K fertilizers while applying N fertilizers; spraying 1% equivalent Bordeaux mixture or 50% carbendazim solution once a month to prevent diseases; after 3 months of subcutaneous sprouting, the paclobutrazol solution with the concentration of 100-150mg/L is applied once.
The graph of the effect of bending branch sprouting is shown in fig. 2. The number of sprouts obtained from each castanopsis hystrix explant after being treated by the above steps is counted, and the results are shown in table 1:
table 1: effect of different substrates on sprouting
The proper matrix can provide better growth environment and required nutrient components, so that the sprouting strips grow more robustly. The implementation effect shows that the number of buds germinated by the original plant taking the mixed soil as the matrix is larger than that of the buds germinated by the original plant taking the loess and the peat soil as the matrix, and the number and the robustness of new buds after scion pruning are also better, which indicates that the mixed soil is more suitable for the branch-pressing and bud-promoting cultivation of the original red cone plant.
Example 2
The present example is substantially the same as the example 1, and the culture medium adopted in the S3 field planting step is mixed soil (the mixed soil is obtained by mixing yellow core soil and peat soil according to a mass ratio of 1: 1), except that in the present example, in the step S2, the red cone branches are trimmed by the following 5 trimming methods, and in the case of using the non-trimmed branches as a Control (CK), 5 groups of the method are set to repeat, and 4 plants are set in each group.
The 5 pruning methods are respectively as follows:
b1: performing top cutting treatment in the same way as in the example 1, cutting off partial side branches on the trunk of the redcone branch, and reserving 2-3 near-top side branches;
b2: performing truncation treatment, and reserving all lateral branches;
b3: without carrying out top cutting treatment, cutting off partial side branches on the trunk of the redcone branch, and reserving 2-3 near-top side branches;
b4: cutting off all side branches on the trunk of the castanopsis hystrix branch, and reserving the trunk and 2-3 near-top leaves;
b5: and (4) cutting off all lateral branches on the trunk of the redcone branch and cutting off the top without leaves.
The final sprouting effect is shown in table 2 and fig. 3.
TABLE 2 Effect of different pruning methods on sprouting Effect
As a result, the mother plants treated by B1, B2 and B3 begin to bud at day 6, and at the moment, 1-4 buds are arranged on 50% of branches, and the part of the buds growing is mostly near the bent branch. By day 11, nearly 80% of shoots had germinated, at which time the maximum number of sprouts was 11, with a maximum of 4.5 cm. By day 18, about 92% of the shoots had germinated, at which time pruning pattern B1 had a maximum of 23 shoots, up to 16 cm. Pruning pattern B4 retained only two or three leaves and also shoots but less fine, pruning pattern B5 did not shoot shoots with no leaves. The control CK had fewer sprouts. As can be seen from the above, the cutting pattern B1 exhibited the best sprouting effect.
Example 3
The operation steps of this example are basically the same as those of example 1, and the culture medium adopted in the planting step of S3 is mixed soil (the mixed soil is obtained by mixing loess and peat soil according to a mass ratio of 1: 1), except that in step S4 of this example, the following 3 branch bending methods are adopted:
a1: the trunk branches are bent by 90-120 degrees and the branches near the top end form a platform;
a2: the trunk branches are bent by 90-120 degrees, and the branches near the top end are pressed to be not higher than the bent parts of the branches;
a3: the trunk branches are bent by 90-120 degrees, and the branches near the top end are pressed down but higher than the bent parts of the branches.
The results are shown in table 3 and fig. 4.
TABLE 3 Effect of the bent shoot mode on sprouting
As a result, the A2 method has good sprouting promoting effect, large sprouting amount and robustness. Furthermore, anova showed that the sprouting effect of treatment a2 was significantly higher than treatments a1 and A3, while the difference between treatments a1 and A3 was not significant. From the above, the germination effect was the best in the A2 treatment method.
The above embodiments are preferred embodiments of the present invention, but the present invention is not limited to the above embodiments, and any other changes, modifications, substitutions, combinations, and simplifications which do not depart from the spirit and principle of the present invention should be construed as equivalents thereof, and all such changes, modifications, substitutions, combinations, and simplifications are intended to be included in the scope of the present invention.
Claims (9)
1. A method for obtaining a large batch of high-quality explants by using a curvicular rosette sprouting promoting technology is characterized by comprising the following steps:
s1, matrix treatment: sterilizing the matrix before field planting, covering with a film, lifting the film before field planting, and watering the matrix thoroughly with water;
s2, trimming: pruning trunk branches of the castanopsis hystrix seedlings to obtain pruned castanopsis hystrix seedlings;
s3, planting: pouring the pruned castanopsis hystrix seedlings obtained in the step S2 into a hole as castanopsis hystrix explant materials, and covering the castanopsis hystrix seedlings with the matrix obtained in the step S1;
s4, bending branches: selecting a part of the pruned trunk of the castanopsis hystrix seedling, which is 5-10 cm away from the base part, as a branch bending part, bending the trunk branch by 90-120 degrees, and fixing the top branch of the pruned castanopsis hystrix seedling;
s5, covering soil: covering the top ends of the branches with a substrate;
s6 sprout management: cutting off lateral sprouting branches until the subcutaneous part begins to sprout and root, and the overground part sprouts into juvenile scion strips; fertilizing, preventing diseases, selectively using a plant growth regulator to obtain a large batch of high-quality explants;
the matrix in the step S1 is obtained by mixing loess and peat soil according to a mass ratio of 1-2: 1;
the trimming processing manner described in step S2 is: cutting off the top, and cutting off the lateral branches to leave 2-3 lateral branches close to the top;
the top shoots described in step S4 are processed as follows: the pressure of the branch near the top end is not higher than that of the branch bending part.
2. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 1, which is characterized in that:
the matrix in the step S1 is obtained by mixing loess and peat soil according to the mass ratio of 1: 1;
the branch bending part in the step S4 is a part, 5cm away from the base, of the trunk of the pruned castanopsis hystrix seedling.
3. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 1, which is characterized in that:
the sterilization described in step S1 is performed using potassium permanganate;
the disinfection in the step S1 is carried out on the day before the permanent planting;
the time for covering the film in the step S1 is 0.5-1.5 days;
watering in the step S1 for 2-3 times;
the fixing in step S4 is performed by using an iron ring or a wooden pile.
4. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 3, wherein the method comprises the following steps:
the sterilization in step S1 is performed by using a potassium permanganate solution with the concentration of 0.5% w/v;
the time for covering the film in step S1 was 1 day.
5. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 1, which is characterized in that:
the castanopsis hystrix seedlings in the step S2 are castanopsis hystrix nutrition bag seedlings;
the inclined placement in the step S3 means that the cut stem of the castanopsis hystrix seedling is placed at an included angle of 5-30 degrees with the ground;
the depth of the cavity in the step S3 is not less than 15 cm;
the substrate coverage depth in the step S3 is 15-25 cm above the base of the trimmed castanopsis hystrix seedling;
the matrix in the step S5 is obtained by mixing yellow core soil, peat soil and pond sludge according to the mass ratio of 1-2: 1: 1.
6. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 5, wherein the method comprises the following steps:
the inclining and placing in the step S3 means that the cut trunk of the castanopsis hystrix seedling is placed at an included angle of 15 degrees with the ground;
the depth of the holes in the step S3 is 15-25 cm;
the substrate coverage depth in the step S3 is 20cm above the base of the trimmed castanopsis hystrix seedling;
the matrix in the step S5 is obtained by mixing yellow core soil, peat soil and pond sludge according to the mass ratio of 1:1: 1.
7. The method for obtaining a large batch of high-quality explants by using the curvicular rosette sprouting promoting technology according to claim 1, which is characterized in that:
the frequency of pruning the lateral shoots of sprouts in the step S6 is 1 time per month;
the specific steps of fertilization described in step S6 are as follows: in 1-3 months after subcutaneous sprouting, applying urea liquid fertilizer 2-3 times per month, and applying P, K fertilizers while applying N fertilizers;
the specific operation for preventing diseases described in step S6 is as follows: spraying 1% equivalent Bordeaux mixture or 50% carbendazim solution once a month to prevent diseases;
the plant growth regulator described in step S6 is paclobutrazol;
the paclobutrazol is used under the condition that a paclobutrazol solution with the concentration of 100-150mg/L is applied once after 3 months of subcutaneous sprouting.
8. The method for obtaining a large batch of high-quality explants by using the curvatura bicolor branch sprouting promoting technology of any one of claims 1 to 7 in the preparation of clonal seedlings of the mangrove bicolor.
9. The application of the method for obtaining a large batch of high-quality explants by the curvatura bicolor branch sprouting promoting technology in the construction of the artificial mangrove bicolor forest according to any one of claims 1 to 7.
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