CN111096173B - Method for promoting pen container tree development through adjusting environmental factors - Google Patents
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Abstract
The invention provides a method for promoting the development of a pen container tree by adjusting environmental factors, in particular to a method for promoting the spore germination and the gametophyte development of the pen container tree by adjusting the matrix type and the porosity of a seedling tray, promoting the seedling growth of the pen container tree after being transplanted by adjusting the matrix type of seedling pots, preliminarily constructing a pen container tree spore breeding system by carrying out factor regulation and control on different picking periods, seeding modes, matrix types, transplanting opportunities and the like, realizing the accurate control seedling breeding of the pen container tree in the whole process of spore, gametophyte, sporophyte and seedling, and effectively solving the technical problem of pen container tree breeding.
Description
Technical Field
The invention relates to the technical field of agricultural cultivation, in particular to a method for promoting pen container tree development by adjusting environmental factors.
Background
The penholder tree (Sphaeropteris lepifera) is tree fern of Cyatheaceae (Cyatheaceae) genus Cyathea (Sphaeropteris), has long trunk, large and dense leaf marks such as umbrella, and is rare fern. The pen container tree not only has excellent ornamental value, but also has important value in the aspects of speciation, ancient climate research, ancient biology and the like, and has important significance in the aspects of researching the relationship between the continental China and the Taiwan plant system. Under natural conditions, the propagation of the pen container trees is mainly carried out by spores generated on leaf backs, but the spores have the defects of short service life, long propagation period and the like, so that the natural growth of the pens is limited; the stems of the pen container trees can be used as culture mediums for orchids, are frequently subjected to destructive felling, and are listed as the first secondary important protection plants in China, because the natural resources are seriously exhausted.
According to investigation, Zhejiang is the north edge of the distribution of the pen container tree, the quantity is extremely rare, the pen container tree germplasm resources in our province are not effectively protected at present, the germplasm resource characteristic characteristics and the information in the aspects of seedling breeding and cultivation are very poor, at present, the technology at the prior stage only simulates the growth conditions of other ferns, researches are carried out on spore germination and gametophyte growth, but the spore germination and gametophyte growth of the pen container tree have the particularity, and the pen container tree is not similar to other common ferns such as Cyathea and the like. According to the research, the pen container tree has strict requirements on the types of the matrixes, namely volume weight, porosity, water retention, air permeability and nutrient supply, the common matrixes such as turfy soil, vermiculite, river sand, ceramsite and other mixed matrixes can not achieve higher spore germination rate, and the matching of the matrixes can not achieve an ideal effect when the subsequent gametophytes grow. In addition, in the prior art, the research on the pen container trees is approximately stopped at the stage of germination and gametophyte breeding, the research on further cultivation of the sporophytes is less, and according to the research, the transplanting survival rate of the pen container tree juvenile sporophyte seedlings is lower, so that the problem of exploring and improving the transplanting survival rate of the pen container tree juvenile sporophyte seedlings is to be solved urgently at present.
In conclusion, based on the technical problem of difficult spore propagation of the pen container trees, the research on the breeding technology of the pen container trees needs to be deeply developed, the proper matrix type suitable for the growth and seedling development of the gametophytes of the pen container trees and the seedling age breeding technology suitable for outplanting are pertinently developed, and the method has important significance for germplasm resource protection and sustainable utilization.
Disclosure of Invention
Based on the technical problems that the spore of the pen container tree has strict requirements on the water retention property, the air permeability and the nutrient supply of a matrix, the spore is very easy to infect germs, pollute and is difficult to breed, the invention provides a method for promoting the development of the pen container tree by adjusting an environmental factor, particularly, the spore germination and the gametophyte development of the pen container tree are promoted by adjusting the matrix type and the porosity of a seedling tray, the matrix type of a seedling pot is adjusted to promote the seedling growth of the transplanted pen container tree in the seedling stage, and a spore breeding system of the pen container tree is preliminarily constructed by carrying out regulation and control on factors such as different picking periods, seeding modes, matrix types, transplanting times and the like, so that the accurate control seedling breeding of the pen container tree in the whole process of the spore, the gametophyte, the sporophyte and the seedling is realized, and the technical problem of breeding of the pen container tree is effectively solved.
In order to achieve the purpose, the invention provides the following technical scheme:
the method for promoting the development of the pen container tree by adjusting the environmental factor comprises the following steps:
(1) regulating the type and porosity of the seedling raising tray substrate to promote spore germination and gametophyte development of the penholder trees;
(2) adjusting the type and porosity of the substrate of the seedling pot to promote the seedling growth of the transplanted pen container trees;
(3) and (3) analyzing the cold resistance and photosynthetic adaptability of the pen container trees at different seedling ages according to low-temperature stress, light response curves, carbon dioxide response curves and fitting data, and making the seedling age of the nursery planting.
Wherein the type and porosity of the matrix of the seedling raising tray in the step (1) are adjusted by adopting sterilized fine peat as the matrix when the seedling raising tray for the pen container trees is used for sowing, and when the screened grain diameter is 3-4mm, the volume weight of the peat matrix is 0.55-0.57g/cm3The total porosity is 70.7-74.4%.
The type of the medium for adjusting the seedling pots in the step (2) is that after 3 months of sowing, when the pen container trees basically form pen container tree sporophytes, the pen container trees are transplanted into the seedling pots, the seedling pots are placed into a greenhouse for cultivation, and ventilation is performed regularly; the substrate of the seedling pot consists of 26-40% of A humus soil, 20-46% of B peat and 27-40% of C sawdust or perlite, wherein the sum of all the components is 100%; the volume weight of the matrix is 0.59-0.69g/cm3The total porosity is between 65 and 69.7%.
Selecting potted seedlings of 2-4 year-old pen container trees with consistent growth vigor, and setting a low-temperature gradient to measure the conductivity and the semi-lethal temperature of the pen container trees with different seedling ages; measuring the light response and carbon dioxide response curves of the pen container trees with different seedling ages by adopting a portable photosynthetic measurement photosynthetic apparatus, and calculating a light saturation point, a light compensation point and CO2Point of compensation and CO2And (5) finding out the seedling age field planting and transplanting of the 3-year-old pen container trees with strong suitability by combining saturation points and transplanting survival rate and growth indexes.
Specifically, the invention also provides an artificial breeding method of the pen container tree, which comprises the following steps:
(1) collecting mature leaves of the pen container tree, and cleaning the leaves until the color of sporangia on the back of the leaves is brown and the sporangia are not completely cracked;
(2) the leaves of the pen container are folded by paper and placed in a shady and dry place for about 4 to 6 days until the sporangium is cracked, and scattered spores are collected and stored at low temperature;
(3) selecting a seedling-raising plate or sowing in an aseptic sowing mode, wherein the sowing density is as follows: 1cm3The spores are sown at 1m2A substrate surface;
(4) growing the gametophyte of the pen container tree about 1.5 months, controlling the temperature and the humidity, and ventilating regularly;
(5) and (3) basically forming pen container tree sporophytes after seeding for 3 months, ventilating at regular intervals, transplanting the pen container trees into seedling pots, putting the seedlings into a greenhouse for cultivation and culturing, and ventilating at regular intervals.
Further, the step (2) is to wrap and fold the leaves of the pen container tree with newspaper or parchment paper and place the leaves in a cool and dry place for about 5 days.
Further, the cryopreservation in the step (2) is to store the collected spores at 4 ℃ or-80 ℃.
Further, in the step (3), when the seedling raising tray is selected for sowing, the substrate is sterilized fine peat with the particle size of 3-4mm, and spores are uniformly sown on the surface after the substrate is thoroughly watered.
Further, the particle size of the fine peat is 4 mm.
Further, in the step (5), when the pen container tree sporophyte seedlings grow to be 6-7cm high, transplanting the pen container tree sporophyte seedlings into a seedling pot for culture.
Furthermore, the substrate of the seedling pot consists of 26-40% of humus soil A, 20-46% of peat B and 27-40% of sawdust C or perlite C, wherein the sum of all the components is 100%.
Furthermore, the substrate of the seedling pot consists of 27% of humus soil A, 46% of peat B and 27% of sawdust C or perlite C.
Further, during the cultivation in the step (4), the temperature is 23-25 degrees, the humidity is about 80-90 percent, the illuminance is 1000-.
Further, during the cultivation in the step (5), the temperature is 20-28 ℃, the humidity needs to be controlled to be above 80%, and the illumination time in the day is 13-14 h/d.
Compared with the prior art, the invention has the beneficial effects that:
1. through a large number of comparative experimental researches, the inventor discovers for the first time that the picking period, the development degree after picking and the seeding density of the spores of the pen container tree obviously influence the growth and the germination rate of the gametophytes. In order to further verify the influencing factors influencing the development of the gametophyte of the pen container tree, L9 (3) is designed4) 3 factor 3 level orthogonal tests, respectively finding out different picking periods, the development degrees (different placing times after picking) after picking and the optimal factor combination of the germination rate of the gametophyte of the superior pencil vase tree under the condition of seeding density: picking leaf with dark brown sporangium on back, wrapping picked leaf with newspaper, placing for 5d, collecting spore, and sowing with sowing density of 1cm3The spores are sown at 1m2Substrate surface (experimental example 5). For germination rate, the reason isThe primary and secondary effects of the element are seeding density>Picking period>Degree of development.
2. Experiments show that the influence of the collection time on the spores of the pen container trees is increased along with the increase of the time after the mature pen container leaves are collected and aired, the naturally falling spores are gradually increased, the spores are basically scattered when the time reaches 5 days, the influence on the formation of gametophytes is small, and after the leaves are placed for 7 days, the spore falling time is too long, a large amount of spores are insufficient in activity or die, and the gametophytes can be formed less.
3. The fertilization of gametophytes is influenced by over-dense or over-sparse inoculation of the spores of the pen container trees, the spores are not favorable for germination, the distribution uniformity of the spores broadcasted by a weighing scoop is poor in the experiment, the gametophytes on the surface of the matrix are intensively developed or part of the surface of the matrix is not grown by the gametophytes, the spores of the pen container trees are placed on paper for finger flick sowing, the sowing uniformity of the pen container trees can be controlled to the maximum degree, and the germination rate is improved.
4. Compared with other ferns, the penholder tree has strict requirements on the substrate, and has better adaptability to mixed substrates and single substrates unlike common Cyathea ferns. The total porosity of the substrate determines the air permeability of the substrate, so that the supply and absorption of water and nutrients and the growth of plants are influenced, and the organic matter content in the substrate is a main source of plant nutrients and can promote the growth and development of the plants. Through field habitat investigation, people pay attention to that the pen container trees are not only wet but also grow in ventilated places, so that in later-period tests, people try to compare various substrates commonly used for ferns, and through a large amount of comparison, people find that the effect of breeding the pen container trees by only adopting peat and one substrate is good, and the effect is better than that of singly using humus, brown soil, vermiculite and river sand or using two or three combined substrates.
5. We find that the pen container tree can realize better germination rate by controlling temperature, humidity and illumination as the conventional Alsophila spinulosa plants, has high requirement on humidity, has high requirements on volume weight and total porosity of matrix, and often has the phenomena of improper matrix and greatly reduced germination rate. Total porosity due to bulk weight of the matrixIndexes influence the water retention and air permeability of the culture medium, and due to the fact that peat has the advantages of being loose and porous, high in water retention, good in air and water permeability, high in water and fertilizer retention and the like, through experimental research, the single medium peat is obtained, and the germination and growth effects are the best after the peat is sieved by 3-4 mm. When the grain diameter of peat is 3-4mm, the volume weight of peat substrate is 0.55-0.57g/cm3The total porosity is 70.7-74.4%, the water retention and air permeability are good, the germination and growth are facilitated, when the screened particle size is less than 2mm, the total porosity of the matrix is reduced, the air permeability and the water permeability of the matrix are relatively poor, and the matrix is susceptible to germs; when the particle size of the sieved particles is larger than 4mm, the volume weight of the matrix is increased, the water retention is reduced and the germination rate is reduced although the air permeability of the matrix is good. Therefore, the spore germination and growth are hindered by the overlarge or undersize peat particles.
6. The requirement of further development of conidia on the type of the matrix is strict, the influence of the peat matrix with the sieving grain size of 4mm on the further development of the sporophyte into the gametophyte is obvious, the green development time of the surface of the matrix is 8d, and the green development time is shorter than other treatment times; and the time of the gametophyte appearing in large quantity and the percentage of the gametophyte developing into the sporophyte also show better growth advantages; through observation under a microscope, we find that the root system of the formed juvenile sporophyte seedling is developed and the root quantity is rich. While the other treated gametophytes grow slightly slower than the peat alone, more gametophytes die during the cultivation process and cannot be fertilized to form sporophytes, i.e., the gametophytes have a low percentage of developing sporophytes.
7. We found that the transplanting timing has an important influence on the transplanting survival rate of the juvenile sporophyte seedlings of the pen container trees, which is different from other Cyathea plants (the transplanting survival rate is higher when the seedling height is smaller and 2-4 cm). When the height of the sporophyte seedlings is less than 5cm, the seedlings are transplanted, the development degree of the root system of the pen container tree is low, the root development of the pen container tree is unfavorable after the seedlings are planted in an excessively small mode, and the transplanting survival rate of the juvenile sporophyte seedlings is low; when the sporophyte seedlings grow to 6-7cm in height (Experimental example 31), the transplanting survival rate is highest after the sporophyte seedlings are transplanted into seedling pots for culture. When the root development of the pen container tree is more than 7cm, the influence of the transplanting on the growth of the seedling revival in the later period of the root is larger, and the survival rate is reduced.
8. The fine peat is favorable for spore germination, but in later growth, the pen container tree has increased demand on substrate nutrients and also likes good permeability, researches show that the substrate is added with some perlite, river sand, ceramsite and the like which are commonly used in the field of original Cyathea plant seedling raising technology, so that the effect on adjusting the water absorption and permeability of the substrate is better, the fine peat has the advantage of high carbon-nitrogen ratio, and the fine peat has a remarkable promoting effect on the seedling growth of the pen container tree. According to the invention, the transplanting time is controlled, the matrix formula is reasonably regulated, the higher transplanting survival rate is obtained, and a technical guidance basis is provided for the field of artificial breeding of the pen container trees. Transplanting when the height of sporophyte seedling is 6-7cm, and the matrix formula A is formed from (wt%) 26-40% of humus soil, 20-46% of peat B and 27-40% of C saw dust or pearlite, in which the sum of all the components is 100%, and the volume weight of matrix is 0.59-0.69g/cm3The total porosity is 65-69.7%, and the transplanting survival rate of the penholder trees is high; and the growth speed is high after 3 months of transplantation.
9. The invention firstly proposes that the cold resistance and photosynthetic adaptability of different seedling ages of the pen container tree are analyzed according to low temperature stress, light response and carbon dioxide response curves and fitting data, and the seedling age of nursery permanent planting is worked out. Zhejiang is the northwest edge of pen container tree distribution, the pen container tree has poor adaptability to low temperature, data are continuously accumulated, the time for outplanting and field planting of the pen container tree is discussed, the cold resistance of the pen container trees with different seedling ages can be known through low-temperature stress, the difference of the light response curves of the pen container trees with different seedling ages is analyzed through photosynthetic property, the rule that the photosynthetic rate of the pen container trees changes along with photosynthetic intensity is reflected through the photosynthetic response curves, and finally CO of the three-year-old pen container tree seedlings is obtained2The compensation point is relatively stable, which indicates that the three-year-old pencil vase tree is in the external CO2At lower concentrations, there is a higher rate of photosynthesis, and thus dry matter can be efficiently accumulated. The invention firstly researches the low-temperature stress and photosynthetic characteristics of different seedling ages to guide production practice, finally obtains the seedling with higher survival rate after outplanting and field planting when the pen container tree grows to three years, the annual average growth rate is higher than that of the two-year and four-year seedlings, and the result is consistent with the photosynthetic characteristic analysis result of the pen container tree, thereby achieving the purpose of combining theory and productionThe effect of (1).
Drawings
FIG. 1 is a drawing showing spore seeding treatment of penholder tree
FIG. 2 is a greenhouse cultivation picture with pen container trees
FIG. 3 is a pen container tree seedling breeding picture
FIG. 4 is a fitting graph of photosynthetic response curves of pen container trees with different seedling ages
FIG. 5 CO of Pen container trees of different seedling ages2Response curve fit plot
FIG. 6 variation of the rate of electrolyte extravasation from leaves of biennial penholder trees after 24h of different low temperature treatments
Detailed Description
The technical solutions of the present invention will be described clearly and completely with reference to the following embodiments of the present invention, and it should be understood that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Example 1
An artificial breeding method of penholder trees comprises the following steps:
(1) collecting mature leaves of the pen container tree, and cleaning the leaves until the color of sporangia on the back of the leaves is brown and the sporangia are not completely cracked;
(2) the leaves of the pen container are folded by paper and placed in a shady and dry place for 5 days until the sporangium is cracked, and scattered spores are collected and stored at low temperature;
(3) selecting a seedling-raising plate or sowing in an aseptic sowing mode, wherein the sowing density is as follows: 1cm3The spores are sown at 1m2A substrate surface;
(4) growing the gametophyte of the pen container tree about 1.5 months, controlling the temperature and the humidity, and ventilating regularly;
(5) and (3) basically forming pen container tree sporophytes after seeding for 3 months, ventilating at regular intervals, transplanting the pen container trees into seedling pots, putting the seedlings into a greenhouse for cultivation and culturing, and ventilating at regular intervals.
Wherein, the step (2) is to wrap and fold the leaves of the pen container trees by newspaper or parchment paper and place the leaves in a cool and dry place for 5 days; the low-temperature preservation of the step (2) is to place the collected spores at 4 ℃; when a seedling raising tray is selected for sowing, the medium is sterilized fine peat with the particle size of 4mm, and spores are uniformly sown on the surface after the medium is thoroughly watered; and (5) transplanting the penholder tree sporophyte seedlings into seedling pots for culture when the penholder tree sporophyte seedlings grow to be 6-7cm high. The substrate of the seedling raising pot consists of 27% of humus soil, 46% of peat and 27% of sawdust; when the cultivation in the step (4) is carried out, the temperature is 25 ℃, the humidity is about 80%, the illuminance is 1000-; and (5) during cultivation, the temperature is 25 ℃, the humidity needs to be controlled at 80%, and the day illumination time is 14 h/d.
Test of the influence of different treatments on spore germination rate of penholder tree
The test method comprises the following steps: sowing tests were carried out at the subtropical crop institute in Zhejiang province between 6 months in 2018 and 6 months in 2019. Collecting fresh mature pen container tree leaves with complete sporangia on the back. Through a plurality of researches of comparison design experiments, the inventor finds that the picking period, the development degree after picking and the seeding density of the spores of the pen container tree obviously influence the growth and the germination rate of the gametophytes. In order to further verify the influencing factors influencing the development of the gametophyte of the pen container tree, L9 (3) is designed4) 3 factor 3 level orthogonal tests, respectively finding out the optimal factor combination of the germination rate of the gametophyte of the superior penholder tree under the conditions of different picking periods, the development degree (different placing time after picking) after picking and the seeding density.
In the specific picking period, pen container tree leaves in different development periods are picked, T1 is that the back sporangium is green, T2 is that the back sporangium is tan and is not completely cracked, and T3 is that the back sporangium is naturally cracked. The specific development degree is that the picked leaves are wrapped by newspaper and then placed for 3d, 5d and 8d respectively, and spores are collected and sown. The specific density is the seeding density: 0.5cm3The spores are sown at 1m21cm at the surface of the substrate3The spores are sown at 1m21.5cm above the substrate surface3The spores are sown at 1m2The surface of the substrate. After sowing, using transparent cover or glass plate to make sealing, and sproutingEnvironmental conditions of the test: the temperature is about 25 degrees, the humidity is about 80 percent, and the illumination is 15-16h every day. The orthogonal test factors and horizontal design are shown in table 1. After 20 days of sowing, the spore germination rate under the microscope field is randomly selected by adopting a sample method, and the average value is repeatedly taken for 3 times.
TABLE 1 orthogonal test factors and horizontal design
Experimental example 1
Picking leaves of pen container tree in different development periods, specifically picking T1 that the spore sac on the back is green, wrapping the picked leaves with newspaper, placing 3d for collecting spore for seeding, wherein the seeding density is 0.5cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 2
Picking leaves of pen container tree in different development stages, specifically picking T1 that the spore sac on the back is green, wrapping the picked leaves with newspaper, placing 5d for collecting spore for seeding, wherein the seeding density is 0.8cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 3
Picking leaves of pen container tree in different development periods, specifically picking T1 that the spore sac on the back is green, wrapping the picked leaves with newspaper, placing 8d for collecting spores for sowing, wherein the sowing density is 1cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 4
Picking leaves of pen container tree in different development periods, specifically picking T2 that the sporangia on the back surface are brown and not completely cracked, wrapping the picked leaves with newspaper, placing 3d for collecting spores for sowing, wherein the sowing density is 0.8cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 5
Picking leaves of pen container tree in different development stages, specifically picking T2 that the sporangia on the back surface is brown and is not completely cracked, wrapping the picked leaves with newspaper, placing 5d for collecting spores for sowing, wherein the sowing density is 1cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 6
Picking leaves of pen container tree in different development stages, specifically picking T2 that the sporangia on the back surface is brown and is not completely cracked, wrapping the picked leaves with newspaper, placing 8d for collecting spores for sowing, wherein the sowing density is 0.5cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 7
Picking leaves of pen container tree in different development periods, specifically picking T3 that sporangia on back naturally crack, wrapping picked leaves with newspaper, placing 3d for collecting spores for sowing, wherein the sowing density is 1cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 8
Picking leaves of pen container tree in different development periods, specifically picking T3 that sporangia on back naturally crack, wrapping picked leaves with newspaper, placing 5d for collecting spores for sowing, wherein the sowing density is 0.5cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
Experimental example 9
Picking leaves of pen container tree in different development periods, specifically picking T3 that sporangia on back naturally crack, wrapping picked leaves with newspaper, placing 8d for collecting spores for sowing, wherein the sowing density is 0.8cm3The spores are sown at 1m2The surface of the substrate. The specific spore collection and seeding method was the same as in example 1.
And (3) test results:
TABLE 2 results of range analysis of orthogonal experiments
As can be seen from Table 2, by setting the test factors, the influence factors and the level of the germination rate of the gametophytes of the pen container trees are obtained. For germination rate, the factor influence is C>A>B, i.e. seeding density>Picking period>The development degree and the preferable parameter combination are A2B2C3, namely, the leaves of the pen container trees in different development stages are picked, specifically, the picked T2 is that the sporangia on the back surface are tan and are not completely cracked, the picked leaves are wrapped by newspaper and placed for 5 days to collect spores for sowing, and the sowing density is 1cm3The spores are sown at 1m2The surface of the substrate. In conclusion, the best combination of the orthogonal experiments is experimental example 5.
Experiments show that the germination rate of the spores of the penholder tree treated by the T1 is low, probably because the sporangium on the back of the penholder tree is green and is immature, the amount of the naturally scattered spores is very small, and the spores basically do not germinate after being sown. In the T2 treatment process, the spores of the penholder trees are mature spores and do not scatter, the spores naturally scatter more after being aired, normal gametophytes can be formed after sowing, the germination rate is high, and the method is the most suitable material for reproducing the spores of the penholder trees. In T3, the sporangium of the pen container tree naturally cracks and scatters, so most of collected spores are sporangium, the number of spores with reproductive capacity is small, and the germination rate of gametophytes after sowing is low.
The influence of the collection time on the spores of the pen container trees is that after the leaves of the mature pen container trees are collected and dried, the naturally falling spores are gradually increased along with the increase of the time, the spores are basically scattered when the leaves reach 5 days, the influence on the formation of gametophytes is small, but after the leaves are placed for 7 days, the spore falling time is too long due to insufficient air humidity, a large amount of spores are insufficient in activity or die, and the gametophytes can be formed less. After the leaves of the penholder tree are collected, the leaves are aired for 4-6 days to collect spores.
The influence of the sowing mode on the spores of the pencil vase trees, the fertilization of gametophytes can be influenced by the over-dense or over-sparse inoculation of the spores of the pencil vase trees, the spores are not favorable for germination, and the distribution uniformity of the spores sowed by a weighing scoop is poor in the experiment, so that the gametophytes on the surface of the matrix develop intensively or no gametophytes grow on part of the surface of the matrixAnd the conidia of the penholder tree are placed on paper for finger flick seeding, so that the seeding uniformity of the penholder tree can be controlled to the maximum degree, and the germination rate is improved. And the seeding density is 1cm3The spores are sown at 1m2The germination rate is highest on the surface of the matrix.
Test the influence of two different matrix types on the propagation and growth of the penholder tree
The test method comprises the following steps: the commonly used substrates for research of pteridophyte include peat, river sand, vermiculite, soil, humus soil, etc. The experiment tries to adopt the mixture ratio of different substrate types as the culture substrate of the pen container trees, namely, the changes of volume weight and total porosity under different substrate formulas and the influence on the propagation condition of the pen container trees are researched. Wherein each treatment had 3 replicates, each replicate 1m2A substrate. After sowing, the germination rate of spores, the time for the matrix surface to appear green, the time for the gametophytes to appear in large numbers and the percentage of the gametophytes to develop into sporophytes in different treatments were recorded. The experimental seeding, germination and other methods and environmental conditions were the same as those of example 1, except for the above-mentioned change in the type of the seeding medium.
The determination method of the volume weight and the porosity of the matrix comprises the steps of taking a container with a known volume (V), weighing a mass (W1), filling the air-dried matrix to be measured, weighing a mass (W2), sealing the container with double layers of gauze (the mass of the gauze is ignored), completely immersing the container filled with the matrix in water for 24 hours, weighing a mass (W3), taking out the container, inverting the container, draining gravity water, weighing a mass (W4), and calculating by using the following formula: volume weight (g/cm)3) (W2-W1)/V; total porosity (%) ═ W3-W2)/V × 100%.
TABLE 3 bulk weight and Total porosity variation for different matrix formulations
And (3) test results:
as can be seen from Table 4, different types of matrices have a greater influence on spore germination and gametophyte development of the penholder trees, which indicates that the penholder trees have strict requirements on the types of matrices. Through long-term comparison, we find that only peat is used for breeding the penholder trees by using a single mediumThe effect is good, and the effect is better than that of singly using humus soil, brown soil, vermiculite and river sand or using two or three combined matrixes. We have also found that the pen container tree has high requirements for humidity, volume weight of the medium, water retention and air permeability, the germination rate of the pen container tree is reduced due to the oversize grain diameter of peat (5 mm of the screened grain diameter of experimental example 13), and the volume weight of the peat medium is 0.55-0.57g/cm when the screened grain diameter is 3-4mm3The total porosity is 70.7-74.4%, the water retention and air permeability are good, the germination rate is high, and the spore germination rate is the highest 93% when the screened particle size is 4 mm; when the particle size of the sieved particles is less than 2mm, the total porosity of the matrix is reduced, the air permeability and the water permeability of the matrix are relatively poor, and the matrix is susceptible to pathogenic bacteria; when the particle size of the sieved particles is larger than 4mm, the volume weight of the matrix is increased, the water retention is reduced and the germination rate is reduced although the air permeability of the matrix is good. Therefore, the spore germination and growth are hindered by the overlarge or undersize peat particles.
In addition, the requirements of the further development of the conidia of the penholder tree on the types of the substrates are strict, and the peat substrate with the sieving grain diameter of 4mm and the volume weight of 0.55g/cm is adopted in the experimental example 123The total porosity is 74.4 percent, the influence on the further development of the sporophyte into the gametophyte is obvious, and the green development time of the matrix surface of the experimental example 12 is 8 days, which is shorter than other treatment times; and the time of the occurrence of the gametophyte in large quantity and the percentage of the gametophyte developing into the sporophyte also show better growth advantages in the 4mm sieved grain size matrix of the experimental example 12, which shows that the growth speed of the gametophyte in the single peat growth matrix is high; through observation under a microscope, the juvenile sporophyte seedlings formed by the treatment of the experimental example 12 are found to have developed root systems and rich root quantities. While the other treated gametophytes grow slightly slower than the peat alone, more gametophytes die during the cultivation process and cannot be fertilized to form sporophytes, i.e., the gametophytes have a low percentage of developing sporophytes.
TABLE 4 influence of different matrix types on spore germination and gametophyte development in penholder trees
Test the influence of three different transplantation periods on the spore propagation of the penholder tree
The test method comprises the following steps: the experimental seedling raising base is arranged in a container seedling raising base in Wenzhou Zhejiang (119-degree 37 '-121-degree 18' E, 2706'-2836' N), and a plurality of layers of shading nets, a spray irrigation and exhaust system and other facilities are arranged inside the experimental seedling raising base, so that the smooth implementation of the experiment is ensured. The test mainly researches the transplanting time of the spore seedlings of the pen container trees and the influence of the difference of transplanting matrixes on the survival rate of transplanting the spore juvenile sporophyte seedlings for 3 months. And (3) selecting different transplanting periods in the test, transplanting the strong pen container tree seedlings which grow well and are consistent in size into a seedling container. The selection of the substrate, humus (passing through a 4mm sieve), peat (passing through a 4mm sieve), sawdust and perlite are used as four variable factors, four levels (0, 1/3, 0.5 and 1.0) are designed, and different light substrate proportions are designed (the substrate is replaced by the substrate shown in the table 6, and other methods are the same as the method in the example 1). The seedling raising container is a non-woven fabric mesh bag (40mm (phi) × 100 mm). Each treatment was performed in 3 replicates, and 20 plants were transplanted in each replicate. The survival rate of transplantation and the height of seedlings after 3 months of transplantation were measured 30 days after transplantation, respectively.
Experimental example 29
The artificial breeding method of the penholder tree is the same as the embodiment 1, and only the transplanting time is replaced: transplanting the sporophyte seedlings to a seedling pot for culture when the sporophyte seedlings grow to be 2-3cm high.
Experimental example 30
The artificial breeding method of the penholder tree is the same as the embodiment 1, and only the transplanting time is replaced: and transplanting the sporophyte seedlings to a seedling pot for culture when the sporophyte seedlings grow to 4-5cm high.
Experimental example 31
The artificial breeding method of the penholder tree is the same as the embodiment 1, and only the transplanting time is replaced: when the sporophyte seedlings grow to 6-7cm high, transplanting the sporophyte seedlings into seedling pots for culture.
Experimental example 32
The artificial breeding method of the penholder tree is the same as the embodiment 1, and only the transplanting time is replaced: and transplanting the sporophyte seedlings to a seedling pot for culture when the sporophyte seedlings grow to be 7-8cm high.
TABLE 5 Effect of different transplantation periods on the survival rate of juvenile sporophyte seedling transplantation
TABLE 6 influence of matrix matching on survival rate of young sporophyte seedling transplantation
And (3) test results:
the transplanting time has important influence on the transplanting survival rate of the pen container tree juvenile sporophyte seedlings, and the test result is different from other Cyathea plants (the transplanting survival rate is higher when the seedling height is smaller). The test results are as follows: when the height of the seedlings is too small, the root development of the pen container tree is not facilitated, and as can be seen from table 5, the seedlings of the sporophyte seedlings are transplanted when the height of the seedlings is less than 5cm, the transplanting survival rate of the juvenile sporophyte seedlings is low, and when the sporophyte seedlings grow to be 6-7cm, the transplanting survival rate is highest after the sporophyte seedlings are transplanted into a seedling pot for cultivation, and is 96.5%. When the root development of the pen container tree is more than 7cm, the influence of the transplanting on the growth of the seedling revival in the later period of the root is larger, and the survival rate is reduced.
As can be seen from the table 6, the fine peat is beneficial to spore germination, but the pen container trees also like good permeability due to increased demand for nutrients of the matrix when growing in the later period. Sporozoite seedlings were grown to 6-7cm height and when the substrate formulation selected Experimental example 47: 27 percent of humus soil, 46 percent of peat and 27 percent of sawdust, and the volume weight is 0.65g/cm3The total porosity is 68.3 percent, and the transplanting survival rate of the penholder trees is the highest, 96.5 percent; and the seedling grows fast after 3 months of transplantation, has the height of 18.6cm, and has obvious advantages compared with other types of substrate proportions. Secondly, the effect of adding perlite into the humus soil and peat mixed matrix is better, such as examples 33 and 40; the requirement of seedling growth on the air permeability of the soil in the later period of the pen container tree is higher, the combination uses humus soil in a single use or two-two combination manner, and the growth effect of peat is better.
Photosynthetic characteristic analysis of pen container trees at four different ages
The test method comprises the following steps: selecting potted seedlings of 2-4 year-old penholder trees with consistent growth vigor, taking 5 complete and 5-11 leaves with no plant diseases and insect pests at the 5 th-11 th leaf position from top to bottom of the current year-old spring and summer tips after precooling exercise for analysis and determination, repeating the steps for 3 times, transferring the leaves into an LF220 computer automatic temperature control cold storage (manufactured by Ningbo Laifu corporation), setting the temperatures to be 4, 0, -2, -4, -6, -8 and-12 ℃ respectively, treating the leaves for 24 hours, and controlling the temperature to be +/-0.11 ℃ (calibrating the leaves by a mercury thermometer), illuminating for 14 hours and keeping the relative humidity at 75%. The conductivity of the sample is measured by referring to the method of Zhangiang (1990), and the semi-lethal temperature (LT50) of the pen container tree is calculated according to the calculation method of the semi-lethal temperature of the related tissues of the Zhugen sea et al (1986) and the Logistic equation.
In addition, 2-4 year-old potted pen container tree seedlings with consistent growth vigor are selected, and the light response and carbon dioxide response curves of pen container trees with different seedling ages are measured by adopting a portable photosynthetic measurement photosynthetic apparatus Li-6400XT (Licor, USA). 6400-02B light source and CO are selected for measuring light response curve2The concentration (Ca) was 400. mu. mol-1The flow rate was 500. mu. mol. s-1The temperature was set at 30 ℃ and the Photosynthetically Active Radiation (PAR) gradient was set at (. mu.mol. m)-2s-1): 1600, 1200, 1000, 800, 600, 400, 200, 150, 100, 75, 50, 25,0. The light source 6400-02B is selected for measuring the carbon dioxide response curve, and the Photosynthetically Active Radiation (PAR) is set to 1200 mu mol.m-2s-1At a flow rate of 500. mu. mol · s-1The temperature was set at 30 ℃ and CO was2Concentration (Ca) gradient was set as: 600, 400,0, 50, 100, 150, 200, 300, 400, 800,1000, 1200. All gas exchange tests were completed at 9:00-11:00 a.m. The data were processed using Excel 2007, SPSS13.0 and Origin photosynthesis software.
TABLE 7 optical compensation points and saturation points of different seedling-age pen container trees
| Variety/index | Optical compensation point (. mu. mol. m)-2s-1) | Light saturation point (. mu. mol. m)-2s-1) |
| Biennial pen container tree | 8.26±2.19b | 194.56±21.45c |
| Three-year-old pen container tree | 6.93±1.86c | 334.56±25.4b |
| Four-year-old pen container tree | 13.2±2.67a | 823.55±30.1a |
TABLE 8 CO of Pen container trees of different seedling ages2Compensation point and saturation point
TABLE 9 influence of different out-nursery planting opportunities on the growth of pen container tree after 1 year of afforestation
And (3) test results:
by researching the dynamic change of the electrolyte extravasation rate of the penholder tree under different low temperature stresses, the electrolyte extravasation rate of the biennial penholder tree leaves shows a change trend (shown in figure 6) which is firstly rapidly increased and then slowly and slowly approximates to an S-shaped curve along with the decrease of low temperature within 24 hours after low temperature treatment, and the electrolyte extravasation rate of the leaves is 155.9-1594.19% under the treatment of the temperature higher than-4 ℃. Under the treatment of minus 10 ℃, the electrolyte extravasation rate of the penholder tree leaves is increased to 4794.19%, and the change trend is gentle when the temperature is as low as minus 10 to minus 12 ℃. The morphologically observed pen container tree freezing condition is consistent with the variation trend of the electrolyte extravasation rate. The electrolyte extravasation rate of the biennial pen container tree leaves is gradually increased to be gentle along with the reduction of the temperature, the electrolyte extravasation rate is approximately increased in an S shape, and the semilethal temperature (LT50) of the biennial pen container tree hand is-8.48 ℃ (Logistic equilibrium y-67.59/(1 +14.33 x exp (0.3139 x))). From the three-year pen container tree, the dynamic change trend of the electrolyte extravasation rate of the pen container tree under different low temperature stresses is slightly gentler than that of the pen container tree for two years, and the electrolyte extravasation rate of the leaves of the three-year pen container tree is reduced by 23.8-34.1% compared with that of the pen container tree for two years under the treatment of the temperature higher than-4 ℃; under the treatment of minus 10 ℃, the electrolyte extravasation rate of the three-year-old penholder tree leaves is reduced by 34.4 to 41.6 percent compared with that of the two-year-old penholder tree leaves. The semilethal temperature (LT50) of the three-year-old pen container tree hand is-10.53 ℃. The low temperature resistance of the seedlings of the three-year-old pencil vase tree is higher.
The light response curves of the pen container trees with different seedling ages are obviously different, and the photosynthetic response curve reflects the rule that the photosynthetic rate of the pen container trees changes along with the photosynthetic intensity. As can be seen from FIG. 4 and Table 7, the net photosynthetic rates of the leaves of the pen container trees at different seedling ages all increased with the increase of the illumination intensity. But when the light saturation point is reached, the net photosynthetic rate growth rate of the pen container tree tends to be flat. However, the optical compensation points and the optical saturation points of the pen container trees with different seedling ages are different obviously, the optical saturation points of the pen container trees with the larger seedling age are higher, the optical compensation points and the optical saturation points of the pen container trees with the four-year age are obviously higher than those of the pen container trees with the other seedling ages, and the optical saturation points of the pen container trees with the four-year age are 4.24 times of those of the pen container trees with the second-year age, which shows that the photosynthetic capacity of the pen container trees is stronger along with the increase of the seedling age.
CO of pen container trees with different seedling ages2The response curves differ significantly, CO2The response curve reflects the net photosynthetic rate of the pen container tree along with CO2The law of concentration change. As can be seen from FIG. 5 and Table 8, the net photosynthetic rates of the leaves of the pencil vase trees at different seedling ages were varied with CO2The concentration increases in an upward direction. But when CO is reached2At the saturation point, the net photosynthetic rate of the pen container tree increases slowly. But CO of pen container trees of different seedling ages2Point of compensation and CO2There is a significant difference in saturation point. CO of biennial seedling-age pen container tree2The compensation and saturation points are significantly higher than other seedling-age pen container trees, of which the CO of three-and four-year-old pen container trees2The saturation point and the compensation point are not significantly different. Shows that the pen container tree grows in the external CO when the seedling age is low (more than 2 years of seedling growth)2The photosynthetic rate is higher under the condition of lower concentration, so that dry matters can be effectively accumulated, and the adaptability is stronger.
In conclusion, the three-year-old pencil vase tree nursery stock CO2The compensation point is relatively stable, which indicates that the three-year-old pencil vase tree is in the external CO2At lower concentrations, there is a higher rate of photosynthesis, and thus dry matter can be efficiently accumulated. When the pen container tree seedlings grow to three years, the survival rate after outplanting and field planting is high and is 84.7%. The average annual growth (57.7 cm for high-year average growth of seedlings and 0.88cm for ground diameter year average growth) is higher than that of biennial seedlings (49.5 cm for high-year average growth of seedlings and 0.65cm for ground diameter year average growth) and that of quadriversal seedlings (54.1 cm for high-year average growth of seedlings and 0.74cm for ground diameter year average growth), which is consistent with the photosynthetic characteristic analysis result of the pen container trees.
Although embodiments of the present invention have been shown and described, it will be appreciated by those skilled in the art that changes, modifications, substitutions and alterations can be made in these embodiments without departing from the principles and spirit of the invention, the scope of which is defined in the appended claims and their equivalents.
Claims (8)
1. The method for promoting pen container tree development by adjusting the environmental factor is characterized by comprising the following steps: (1) regulating the type and porosity of the seedling raising tray substrate to promote spore germination and gametophyte development of the penholder trees; (2) adjusting the type and porosity of the substrate of the seedling pot to promote the seedling growth of the transplanted pen container trees; (3) according to low-temperature stress, light response and carbon dioxide response curves and fitting data, analyzing cold resistance and photosynthetic adaptability of different seedling ages of the pen container trees, and making the seedling age of nursery permanent planting;
wherein the type and porosity of the matrix of the seedling raising tray in the step (1) are adjusted by adopting sterilized fine peat as the matrix when the seedling raising tray of the pen container tree is used for sowing, the sieving particle size is 3-4mm, and the volume weight of the peat matrix is 0.55-0.57g/cm3Total porosity 70.7-74.4%;
the type of the medium for adjusting the seedling pots in the step (2) is that after 3 months of sowing, when the pen container trees basically form pen container tree sporophytes, the pen container trees are transplanted into the seedling pots, the seedling pots are placed into a greenhouse for cultivation, and ventilation is performed regularly; the substrate of the seedling pot consists of 26-40% of A humus soil, 20-46% of B peat and 27-40% of C sawdust or perlite, wherein the sum of all the components is 100%; the volume weight of the matrix is 0.59-0.69g/cm3And the total porosity is 65-69.7%.
2. The method of claim 1, wherein the substrate of the pot for raising seedlings is composed of 27% of humus A, 46% of peat B, 27% of sawdust C or perlite C.
3. The method according to claim 1, wherein the step (2) is transplanting the penholder tree sporophyte seedlings to a nursery pot for cultivation when the seedlings grow to 6 to 7cm in height.
4. The method as claimed in claim 1, wherein the step (3) is to select potted seedlings of 2-4 year old pen container trees with consistent growth vigor, and to set a low temperature gradient to measure the conductivity and semi-lethal temperature of the pen container trees with different seedling agesDegree; measuring the light response and carbon dioxide response curves of the pen container trees with different seedling ages by adopting a portable photosynthetic measurement photosynthetic apparatus, and calculating a light saturation point, a light compensation point and CO2Point of compensation and CO2And (5) finding out the planting and transplanting of the 3-year pen container seedling age with strong suitability by combining the saturation point with the transplanting survival rate and the growth index.
5. The method as claimed in claim 1, wherein the incubation in step (1) is performed at a temperature of 23-25 ℃, a humidity of 80% -90%, an illumination intensity of 1000 and 1500lx, and a time of 15-16 h/d.
6. The method of claim 1, wherein the cultivation in step (2) is carried out at a temperature of 20-28 ℃, a humidity of 80% or more and a day illumination time of 13-14 h/d.
7. The method as claimed in claim 1, further comprising a seed collection step before the sowing of the penholder tree, specifically collecting mature penholder tree leaves, and cleaning the leaves until the color of sporangia on the back of the leaves is brown and the spores are not completely cracked; the leaves of the pen container are folded by paper and placed in a shady and dry place for 4 to 6 days until the sporangium is cracked, and scattered spores are collected and stored at low temperature; selecting a seedling-raising plate or sowing in an aseptic sowing mode, wherein the sowing density is as follows: 1cm3The spores are sown at 1m2A substrate surface; growing the gametophyte of the pen container tree in 1.5 months, controlling the temperature and the humidity and ventilating regularly.
8. The method of claim 7, wherein the leaves of the pen container tree are wrapped with newspaper or parchment paper and placed in a cool and dry place for 5 days.
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Address after: University Science Park, No.38 Dongfang Road, Ouhai District, Wenzhou City, Zhejiang Province Applicant after: Zhejiang Subtropical Crops Research Institute Address before: 325000 Zhejiang Subtropical Crop Research Institute, No. 334, Xueshan Road, Ouhai District, Wenzhou City, Zhejiang Province Applicant before: Zhejiang Subtropical Crops Research Institute |
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