CN115669402A - Method and equipment for high-temperature restoration of fertility of lily distant hybrid - Google Patents
Method and equipment for high-temperature restoration of fertility of lily distant hybrid Download PDFInfo
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Abstract
The invention provides a method and equipment for restoring fertility of lily distant hybrids at high temperature, wherein the method for restoring fertility of the lily distant hybrids at high temperature comprises the following steps: selecting buds which are in a meiosis prophase I-metaphase I from lily buds as buds to be treated; respectively determining the shape and the size of the heating bag according to the character characteristics of each flower bud to be treated; installing each heating bag on the corresponding flower bud to be treated; and sequentially carrying out high-temperature heating treatment on the flower buds to be treated according to the preset heating temperature and heating time to obtain the fertile pollen. The method and the equipment for restoring the fertility of the lily distant hybrid provided by the invention aim to solve the problems of sterility of the lily distant hybrid and long breeding period, and solve the problems that the whole development process of plants is influenced by high temperature and the plants with high body size are difficult to process by targeted processing.
Description
Technical Field
The invention relates to the technical field of plant breeding, in particular to a method and equipment for restoring fertility of lily distant hybrids at high temperature.
Background
Lily belongs to Liliaceae (Liliaceae) Lilium plants, is an important bulbous flower in the world, has large and bright color and unique flower posture, is one of world famous flowers, is widely applied to gardens, pot flowers, particularly fresh cut flowers and the like, and is deeply favored. In China, lily belongs to high-grade cut flowers, the sale price is obviously higher than that of other cut flower varieties, and in recent years, LA and OT type lily obtained by distant hybridization of musk lily and Asian lily, trumpet lily and oriental lily is deeply favored by the market due to high resistance and rich color.
Therefore, interspecific distant hybridization is an important means and breeding trend for breeding new lily varieties, and has the advantage that valuable horticultural traits from different lily genomes can be combined or introduced into hybrid varieties by combining the horticultural characteristics of Asian lily, musk lily, oriental lily and trumpet lily. However, most distant hybrids show high sterility due to the large genetic distance, and the chromosomes cannot be normally paired and separated during meiosis, and are difficult to be used for further improvement and introgression breeding of lilies. Therefore, how to restore the fertility of the distant hybrid becomes a bottleneck problem for cultivating a new variety of excellent lily distant hybrids.
Disclosure of Invention
The invention provides a method and equipment for restoring fertility of a lily distant hybrid at high temperature, which aims to solve the problems of sterility and long breeding period of the lily distant hybrid in the traditional technology and solve the problems of influence on the whole development process of a plant and high plant size caused by high temperature through targeted treatment.
In a first aspect, the present invention provides a method for restoring fertility to a lily distant hybrid at a high temperature, comprising:
selecting buds which are in a meiosis prophase I-metaphase I from lily buds as buds to be treated;
respectively determining the shape and the size of the heating bag according to the character characteristics of each flower bud to be treated;
installing each heating bag on the corresponding flower bud to be treated;
and sequentially carrying out high-temperature heating treatment on the buds to be treated according to the preset heating temperature and heating time to obtain the fertile pollen.
Further, before selecting the bud at meiosis prophase i-metaphase i in the lily bud as the bud to be treated, the method further comprises the following steps: establishing a corresponding relation between the development stage of microsporocyte of the lily buds and the external morphological characteristics of the lily buds so as to select the buds at early stage I-middle stage I of meiosis according to the shape of the buds.
Further, after each of the heating bags is mounted on the corresponding bud to be treated, the method further comprises the following steps: and sealing the heating bag to form a closed space in the heating bag.
Further, after sealing the heating bag to form a sealed space inside the heating bag, the method further comprises: support rods and a wire binding belt are arranged on the lily branches to fix the connecting circuits of the heating bags.
Furthermore, the heating temperature of each bud to be treated is 40-44 ℃, and the heating time is 4-6 h.
The present invention further provides an apparatus for restoring fertility of a lily distant hybrid at high temperature, comprising:
the heating bag comprises a plurality of heating bags, wherein a heating cavity is arranged in each heating bag, and a heater and a temperature sensor are arranged in each heating cavity;
the temperature controller is respectively electrically connected with each heater and each temperature sensor and is used for controlling the heating temperature in each heating bag; and (c) a second step of,
and the battery is electrically connected with the controller, the heaters and the temperature sensors.
Furthermore, a heat insulation layer and a fireproof heat insulation layer are sequentially arranged in the heating bag.
Furthermore, the heating element is set as a heating sheet, the heating sheet is arranged on the heat insulation layer, and the temperature sensor is arranged on the heating sheet.
Furthermore, each heating bag is provided with an opening communicated with the heating cavity, and each opening is provided with a sealing strip used for sealing the opening.
Further, the battery is also electrically connected with an optical coupling isolation relay.
The method for restoring the fertility of the lily distant hybrid at high temperature provided by the invention can be adapted to lily buds with different sizes and growth forms by arranging the heating bags with different shapes and sizes, can pertinently select the lily buds corresponding to the growth and development stages for heating treatment, and has the advantages of simple operation, small toxicity, low cost, low bud falling rate and higher success rate of restoring the fertility of lily for herbaceous plants such as lily with inconsistent bud development and weak stems. According to the invention, lily buds are heated and induced to generate fertile pollen of which the concentration is up to 36.64%, so that fertility restoration is realized, and the lily buds can be directly used for cross breeding and pollination or used for breeding backcross first-generation hybrid; in addition, the method for restoring the fertility of the lily distant hybrid at high temperature provided by the invention can further shorten the breeding time of the backcross first generation of the lily on the basis of restoring the fertility of the lily, promote the gradual infiltration and mutation of the genes of the lily and enrich the genetic composition of the lily.
Drawings
In order to more clearly illustrate the technical solutions of the present invention or the prior art, the drawings needed for the description of the embodiments or the prior art will be briefly described below, and it is obvious that the drawings in the following description are some embodiments of the present invention, and those skilled in the art can also obtain other drawings according to the drawings without creative efforts.
FIG. 1 is a schematic flow chart of a method for restoring fertility to a lily distant hybrid at a high temperature according to the present invention;
FIG. 2 is a photomicrograph of pollen from example one (red is pollen that has been treated to restore fertility, and the remaining dark brown is sterile pollen);
FIG. 3 is a photomicrograph of pollen from example two;
FIG. 4 is a photomicrograph of pollen from example III;
FIG. 5 is a photomicrograph of pollen from example V;
FIG. 6 is a photomicrograph of pollen in comparative example I;
FIG. 7 is a schematic structural diagram of an apparatus for restoring fertility to lily distant hybrids at high temperature according to the present invention;
FIG. 8 is a schematic structural elevation view of the heating pouch of FIG. 7;
FIG. 9 is a schematic cross-sectional structural view of the heating pouch of FIG. 7;
FIG. 10 is a schematic view of a portion of the structure of FIG. 7;
reference numerals: 1: equipment for restoring fertility of lily distant hybrids at high temperature; 2: heating the bag; 3: a heating cavity; 4: a heating plate; 5: a temperature sensor; 6: a heat-insulating layer; 7: a thermal insulation layer; 8: a battery; 9: a temperature controller; 10: multichannel opto-isolator relay.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention clearer, the technical solutions of the present invention will be clearly and completely described below with reference to the accompanying drawings, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. 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.
In the description of the embodiments of the present invention, it should be noted that the terms "central", "longitudinal", "lateral", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc. indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience of description and simplicity of description, but do not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and thus, should not be construed as limiting the embodiments of the present invention. Furthermore, the terms "first," "second," and "third" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance.
In the description of the embodiments of the present invention, it should be noted that, unless explicitly stated or limited otherwise, the terms "connected" and "connected" are to be interpreted broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; may be directly connected or indirectly connected through an intermediate. Specific meanings of the above terms in the embodiments of the present invention may be understood as specific cases by those of ordinary skill in the art.
In embodiments of the invention, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may be directly contacting the first and second features or indirectly contacting the first and second features through intervening media. Also, a first feature "on," "over," and "above" a second feature may be directly or diagonally above the second feature, or may simply indicate that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature may be directly under or obliquely under the first feature, or may simply mean that the first feature is at a lesser elevation than the second feature.
In the description herein, references to the description of the term "one embodiment," "some embodiments," "an example," "a specific example," or "some examples," etc., mean that a particular feature, structure, material, or characteristic described in connection with the embodiment or example is included in at least one embodiment or example of an embodiment of the invention. In this specification, the schematic representations of the terms used above are not necessarily intended to refer to the same embodiment or example. Furthermore, the particular features, structures, materials, or characteristics described may be combined in any suitable manner in any one or more embodiments or examples. Moreover, various embodiments or examples and features of various embodiments or examples described in this specification can be combined and combined by one skilled in the art without being mutually inconsistent.
The method and apparatus for restoring fertility to lily distant hybrid at high temperature according to the present invention will be described with reference to FIGS. 1 to 10.
First, the present invention provides a method for restoring fertility to a lily distant hybrid at a high temperature, comprising:
s10, selecting buds in the meiosis prophase I-metaphase I from the lily buds as buds to be treated;
s20, respectively determining the shape and the size of the heating bag according to the character characteristics of each flower bud to be treated;
s30, mounting each heating bag on the corresponding flower bud to be treated;
and S40, sequentially carrying out high-temperature heating treatment on the flower buds to be treated according to preset heating temperature and heating time to obtain fertile pollen.
For most distant hybrids, a high degree of sterility is exhibited due to the inability of chromosomes to pair and segregate normally during meiosis, limiting further improvement and introgression breeding of lilies. Therefore, how to restore fertility of distant hybrids becomes a bottleneck problem in breeding new varieties of excellent lily distant hybrids. In view of the above, the method for restoring fertility of the lily distant hybrid at high temperature provided by the invention can restore fertility of the lily distant hybrid by treating lily buds at high temperature.
Specifically, at the initial stage of the experiment, it is first required to determine the flower buds to be treated on the lily, and the flower buds of the lily at prophase i-metaphase i are selected as the flower buds to be treated, specifically, whether microsporocyte is at prophase i-metaphase i is observed by collecting anthers in the flower buds and by sheet production, which is not limited by the invention.
The traditional high-temperature treatment method for the herb flowers generally places the whole plant of the herb flowers in a heating greenhouse or an incubator, however, the herb flowers are heated unevenly at various parts, and the herb flowers may die during the long-time heating process. Furthermore, because the lily buds need to be heated at high temperature, in the method provided by the invention, respective treatment means are provided for the condition that the development stages of the lily buds are inconsistent, and heating components such as heating bags are provided for the characteristics that the branches of the lily buds are soft and the buds are tender, so that the heating bags are light and convenient to install on the buds, and the buds are prevented from being damaged. And because the shapes and the sizes of the lily buds are different, different heating bags can be arranged to be matched with the lily buds according to the characteristics of the buds to be treated, so that the buds can be strictly wrapped in the heating bags.
In the technical scheme provided by the invention, each bud to be processed of the lily can be respectively heated through the heating bag, and the heating temperature and the heating time can be respectively controlled, so that the targeted treatment of different buds to be processed is facilitated, the whole method is simple in flow and high in success rate, and the efficiency of the high-temperature fertility restoration treatment of the lily and the success rate of experiments can be remarkably improved; in addition, it should be noted that, for the flower buds to be treated in the early stage i-middle stage i of meiosis, different heating temperatures and heating times can be set for different flower buds to be treated to form a treated temperature gradient and a treated time gradient, so as to obtain the optimal experimental conditions in the temperature gradient and the time gradient according to the experimental effect.
According to the method for recovering the fertility of the lily distant hybrid at the high temperature, provided by the invention, through arranging the heating bags with different shapes and sizes, the method can be adapted to lily buds with different sizes and growth forms, can pertinently select the lily buds corresponding to the growth and development stages for heating treatment, and is simple to operate, low in toxicity, low in cost, low in bud falling rate and high in success rate of recovering the fertility of lily and other herbal plants with inconsistent bud development and weak stems. The invention induces the lily buds to generate fertile pollen with the rate of 36.64 percent by heating the lily buds, recovers the fertility, and can be directly used for hybrid breeding pollination or used for breeding backcross first-generation hybrid; in addition, the method for restoring the fertility of the lily distant hybrid at high temperature provided by the invention can further shorten the breeding time of the backcross first generation of the lily on the basis of restoring the fertility of the lily, promote the gradual infiltration and mutation of the genes of the lily and enrich the genetic composition of the lily.
Further, before S10, selecting a bud at meiosis early stage i-middle stage i in lily buds as a bud to be treated, the method further comprises: s100, establishing a corresponding relation between the microsporocyte development stage of the lily buds and the external morphological characteristics of the lily buds so as to select the buds at prophase I-metaphase I of meiosis according to the shape of the buds.
In the technical scheme provided by the invention, the flower buds in the prophase I-metaphase I of meiosis can be selected according to the shape of the flower buds by establishing the corresponding relation between the development stage of microsporocyte mother cells of the flower buds and the external morphological characteristics of the flower buds of lily. Specifically, a plurality of representative anthers in the flower buds with different shapes can be collected, and the microsporocyte development stage of the anthers can be observed through flaking, so that the corresponding relation between the microsporocyte development stage of the lily flower buds and the external morphological characteristics of the lily flower buds can be statistically established. Please refer to the table below, which is a comparison table of the common bud length, anther length and corresponding microsporocyte development stage of lily.
TABLE 1 correlation of OT lily 'Black Beauty' bud length with the meiotic progression of microsporocytes
Further, after S40, installing each of the heating bags on the corresponding bud to be treated, the method further includes: s200, sealing the heating bag to form a closed space in the heating bag. In order to prevent heat loss and form a relatively closed space in the heating bag, in the technical scheme provided by the invention, the heating bag is subjected to sealing treatment, specifically, the sealing bag can be bound on the branches of the lilies by using a binding tape to complete sealing of the sealing bag, or a sealing strip or a sealing adhesive sheet can be arranged at the opening of the sealing bag to complete sealing of the sealing bag, which is not limited by the invention.
Furthermore, as the heating bag needs to be heated, more connecting lines exist, so as to ensure the stability of the connection of the sealing bag and the lily branches and prevent the branches from being crushed or bent. In the technical scheme provided by the invention, after S40, sealing the heating bag to form a closed space in the heating bag, the method further comprises the following steps: s300, arranging support rods and a wire binding belt on the lily branches to fix connecting lines of the heating bags. The support rods can be arranged on the main branches of the lilies, the wire harness belts can bundle up the wire harnesses to prevent the wire harnesses from being scattered and fix the wires, and the branches are dispersed to bear weight on the support rods, so that the stable installation effect can be ensured.
Specifically, the present invention treats OT lily (hybrid progeny of oriental lily and trumpet lily) 'Black Beauty' using high temperature, and provides several examples as follows:
example 1:
determining the flower buds to be treated: using OT lily 'Black Beauty' young flower buds as test materials, collecting pollen tablets in flower buds with different lengths, observing a meiosis process, and selecting the flower buds in the early stage I-middle stage I of meiosis according to the relationship between the length of the flower buds and the meiosis process of microsporocyte meiosis. Heating the flower buds to be treated by using a heating bag: the buds are treated by continuously heating for 4 hours at 40 ℃.
Example 2:
the difference from example 1 is that the flower buds are treated by continuous heating at 40 ℃ for 6h.
Example 3:
the difference from example 1 is that the buds are treated by continuous heating at 42 ℃ for 4 h.
Example 4:
the difference from example 1 is that the flower buds are treated by continuous heating at 42 ℃ for 6h.
Example 5:
the difference from example 1 is that the flower buds are treated by continuous heating at 44 ℃ for 4 h.
Example 6:
the difference from example 1 is that the flower buds are treated by continuous heating at 44 ℃ for 6h.
Comparative example 1:
the flower buds were not subjected to heat treatment.
When the flower buds of examples 1-6 and comparative example 1 are developed in the following process, counting the flower bud falling rate, the flower bud falling rate (%) = the number of dead and fallen flower buds/the total number of treated flower buds multiplied by 100, collecting the anther of the pollen to be scattered, scattering the pollen in a cool and ventilated place, flaking the dried pollen and observing the fertility of the treated pollen. Microscopic observation shows that OT lily 'Black Beauty' is a distant hybrid, the pollen fertility is 0, and the pollen successfully induced at high temperature has vigor and can be used for pollination or breeding backcross first generation.
Table 2: percentage of active pollen and percentage of bud falling for examples 1-6 and comparative example 1
As can be seen from the above table, the optimal heating temperature is 42 ℃, the optimal heating time is 4h, the induction rate of the pollen with activity can reach 36.64 percent at most, the bud falling rate is 40 percent, and all the treated buds of the combination can generate the pollen with activity.
On the basis of the method for restoring the fertility of the distant hybrid lily at high temperature, the invention also provides equipment 1 for restoring the fertility of the distant hybrid lily at high temperature, which comprises the following steps: a heating bag 2, a temperature sensor 5 and a battery 8. The heating bags 2 comprise a plurality of heating bags 2, and the plurality of heating bags 2 respectively correspond to a plurality of buds of the lilies on the same single plant or different single plants at the same development stage. The heating chambers 3 are arranged in the heating bags 2, and the heating chambers 3 can be adjusted to be cylinders, cuboids or shapes matched with the flower buds according to the shape and the size of the flower buds, and the invention is not limited to this. The heating bag 2 is sleeved outside the flower buds, a heater and a temperature sensor 5 are arranged in the heating cavity 3, the flower buds are heated at constant temperature by the whole heating cavity, the heater provides a heating function, and constant temperature adjustment is realized by matching with the temperature sensor, so that overhigh or overlow temperature is prevented; the temperature controller 9 is respectively electrically connected with the heater and the temperature sensor 5 and is used for controlling the heating temperature in the heating bags 2 so as to ensure that each heating bag 2 heats the flower buds according to the preset temperature; the battery 8 is electrically connected with the temperature controller 9, the heater and the temperature sensor 5, and in the technical scheme provided by the invention, the battery 8 is preferably a 12V outdoor lithium battery 8.
Specifically, in order to ensure a good heating effect, a heat insulation layer 6 and a fireproof heat insulation layer 7 are arranged in the heating bag 2. In this embodiment, the fireproof layer is a fireproof cotton heat-insulating layer with aluminum foil, and the heat-insulating layer 6 is a coated kraft heat-insulating layer. Further, the heating member is set as a heating sheet 4, the heating sheet 4 is set on the heat-insulating layer 6, and the temperature sensor 5 is set on the heating sheet 4 for sensing the heating temperature conveniently.
Furthermore, the heating bag 2 is provided with openings communicated with the heating cavity 3, and each opening is provided with a binding belt, so that the openings can be tightly bound by the binding belts after the flower buds are sleeved with the heating bag 2, the temperature loss is prevented, and a relatively closed space is formed in the bag. In addition, battery 8 still electricity is connected with opto-coupler isolation relay 10 for whole equipment is safer, portable, and improves duration. It should be noted that, because the device has a plurality of lines corresponding to different heating bags 2 and flower buds, the electric wires and lily branches can be bound by using the wire binding belt and the support rods in the specific use process, so that the stability of connection between the device and lily is ensured.
The equipment 1 for restoring the fertility of the lily distant hybrid at high temperature provided by the invention not only can be used for a method for restoring the fertility of the lily distant hybrid at high temperature, but also can be used for treating large herbaceous flower plants which are soft in branches, unbalanced in bud development stage and difficult to place in a constant temperature box at high temperature. The equipment can realize bagging of one flower bud, and solves the problem of unbalanced development stage of the flower bud; the device uses a 12V outdoor lithium battery, so that the device is safer and more portable, and has better cruising ability; because the equipment is provided with the temperature controller 9, a plurality of buds can be processed at one time, the efficiency is higher, the temperature of each heating bag 2 can be accurately adjusted and controlled, and the requirements of each experimental combination are met; in addition, the heating bag 2 that this equipment set up is light in weight, and cooperation brace rod and bunch area can avoid the flowering branch rupture, effectively ensure the treatment effect.
Finally, it should be noted that: the above examples are only intended to illustrate the technical solution of the present invention, and not to limit it; although the present invention has been described in detail with reference to the foregoing embodiments, it should be understood by those of ordinary skill in the art that: the technical solutions described in the foregoing embodiments may still be modified, or some technical features may be equivalently replaced; and such modifications or substitutions do not depart from the spirit and scope of the corresponding technical solutions of the embodiments of the present invention.
Claims (10)
1. A method for restoring fertility of a lily distant hybrid at a high temperature, comprising:
selecting buds which are in a meiosis prophase I-metaphase I from lily buds as buds to be treated;
respectively determining the shape and the size of the heating bag according to the character characteristics of each flower bud to be treated;
installing each heating bag on the corresponding flower bud to be treated;
and sequentially carrying out high-temperature heating treatment on the buds to be treated according to the preset heating temperature and heating time to obtain the fertile pollen.
2. The method according to claim 1, wherein before selecting the buds at pre-meiosis stage i-mid stage i in lily buds as the buds to be treated, the method further comprises: establishing a corresponding relation between the development stage of microsporocyte of the lily buds and the external morphological characteristics of the lily buds so as to select the buds at early stage I-middle stage I of meiosis according to the shape of the buds.
3. The method of claim 1, further comprising, after mounting each of the heating bags on the corresponding flower bud to be treated: and sealing the heating bag to form a closed space in the heating bag.
4. The method of claim 1, further comprising, after sealing the heater bag to form a sealed space within the heater bag: support rods and a wire binding belt are arranged on the lily branches to fix the connecting circuits of the heating bags.
5. The method according to claim 1, wherein each of the flower buds to be treated is heated at a temperature of 40 ℃ to 44 ℃ for a time of 4h to 6h.
6. An apparatus for restoring fertility to a lily distant hybrid at a high temperature, comprising:
the heating device comprises a plurality of heating bags, wherein a heating cavity is arranged in each heating bag, and a heater and a temperature sensor are arranged in each heating cavity;
the temperature controller is respectively electrically connected with each heater and each temperature sensor and is used for controlling the heating temperature in each heating bag; and the number of the first and second groups,
and the battery is electrically connected with the controller, the heaters and the temperature sensors.
7. The apparatus for restoring fertility to a lily distant hybrid at a high temperature according to claim 6, wherein a heat insulating layer and a fireproof heat insulating layer are sequentially disposed in the heating bag.
8. The apparatus for restoring fertility to a lily distant hybrid at a high temperature according to claim 7, wherein the heating member is a heating sheet, the heating sheet is disposed on the heat-insulating layer, and the temperature sensor is disposed on the heating sheet.
9. The apparatus for restoring fertility to a lily distant hybrid at high temperature according to claim 6, wherein each heating bag is provided with an opening communicating with the heating cavity, and each opening is provided with a sealing strip for closing the opening.
10. The apparatus for high temperature restoration of lily distant hybrid fertility according to claim 6, wherein said battery is further electrically connected with an opto-isolator relay.
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