CN115316252B - Method for establishing root division system in herbaceous root hemiparasitic plant vessel - Google Patents
Method for establishing root division system in herbaceous root hemiparasitic plant vessel Download PDFInfo
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Classifications
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
- A01G31/02—Special apparatus therefor
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P60/00—Technologies relating to agriculture, livestock or agroalimentary industries
- Y02P60/20—Reduction of greenhouse gas [GHG] emissions in agriculture, e.g. CO2
- Y02P60/21—Dinitrogen oxide [N2O], e.g. using aquaponics, hydroponics or efficiency measures
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- Life Sciences & Earth Sciences (AREA)
- Environmental Sciences (AREA)
- Biodiversity & Conservation Biology (AREA)
- Ecology (AREA)
- Forests & Forestry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Botany (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
- Cultivation Of Plants (AREA)
Abstract
The invention relates to a method for establishing a root dividing system in a herbaceous root hemiparasitic plant dish, relates to the technical field of culture in a plant dish, and aims to solve the problem that an effective root dividing system is lacked in the root behavior research process of root hemiparasitic plants. On the basis of fully understanding the root development characteristics of the half-parasitic plants at the root of the herbaceous plant, the invention obtains an ideal rooting material by selecting conditions such as reasonable seedling types, root cutting positions, culture time and the like, and establishes an effective in-vessel rooting system by utilizing the physical barrier of the two-grid culture vessel. The method solves the problem of uneven distribution of the root systems at two sides of the root system in the root dividing process of the herb root hemiparasitic plant, improves the accuracy and reliability of research, and provides a convenient method and a convenient system for researching the root system behavior of the herb root hemiparasitic plant seedling.
Description
Technical Field
The invention relates to the technical field of botany, in particular to a method for establishing a root dividing system in a herbaceous root hemiparasitic plant vessel.
Background
The root hemiparasitic plant is a parasitic plant which has a green leaf and retains a certain photosynthetic capacity, but a parasitic organ (an aspirator) is formed at the root to obtain resources such as nutrients from a host. Root hemiparasitic plants are the main group of parasitic plants, more than half of the parasitic plants worldwide are root hemiparasitic plants, and in our country this proportion is up to 75%. Some root hemiparasitic plants are important resource plants, but some species are also malignant weeds, and have great ecological and economic impact. However, due to lack of systematic understanding of the parasitic processes and control mechanisms of root hemiparasitic plants, both parasitic weed control and resource utilization of root hemiparasitic plants are passive. Since the parasitic process of the root hemiparasitic plant occurs underground, the root behavior directly affects the establishment of the parasitic relationship and the parasitic intensity. Therefore, the research on the root system behaviors of the root hemiparasitic plants, including response processes and related regulation and control mechanisms of the root system to various biological and non-biological environmental factors, is the basis for analyzing the establishment and regulation and control processes of the parasitic relationship between the root system and the host, and can provide scientific guidance for the high-efficiency prevention and control of the parasitic weeds and the reasonable utilization of the resource root hemiparasitic plants.
In the research of plant root system behaviors, the root division system is widely applied. The root system is arranged the root system in different compartments respectively according to the experiment demand, has physical isolation between each compartment, carries out independent processing to the root system of different parts under the condition of sharing same aboveground part to the response of root system to heterogeneous soil environment under the simulation research natural growth state. As early as the 40's of the 20 th century, the rooting system was used in plant metabolism and development research, and plays an important role, especially in distinguishing whether a response is affected locally or is regulated systemically. Most plants with fibrous root systems are easier to root, but for plants with fibrous root systems, the establishment of root systems is relatively difficult, and especially for experimental treatments requiring more uniform root division, the root division system is more difficult to control. Root hemiparasitic plants are plants with a straight root system, most of the plants are difficult to independently survive, the root system grows slowly, and uniform root division is difficult. The suction device is a characteristic organ of the root hemiparasitic plant and is also an important focus in the research of root system behaviors of the root hemiparasitic plant. Therefore, in the process of establishing the root division system of the root hemiparasitic plant, in addition to ensuring uniform root division, the capacity of generating the suction device of the root system is also very critical. Until now, no report of successfully establishing root system of root hemiparasitic plant exists, which results in limited research on root system behavior of root hemiparasitic plant.
In order to solve the problem that an effective root system is lacked in root system behavior research of root hemiparasitic plants, deep and systematic research needs to be carried out aiming at the root system development characteristics of the root hemiparasitic plants, and a root system establishment method suitable for the root system development characteristics of the root hemiparasitic plants is provided on the basis. The method has important significance for researching the root system behavior and the response mechanism of the root hemiparasitic plant.
Disclosure of Invention
In order to solve the problems in the related art, the invention provides a method for establishing a root dividing system in a herbaceous root hemiparasitic plant vessel. The invention obtains healthy and strong aseptic seedlings by optimizing seeds with good maturity, reasonably storing and sterilizing the seeds and promoting germination in time, obtains ideal root-dividing materials by selecting conditions such as reasonable seedling types, root-cutting positions, culture time and the like, establishes an effective in-vessel root-dividing system and solves the technical problem that an effective root-dividing system of half-parasitic plants on herb roots is lacked at present.
The invention provides a method for establishing a root division system in a herbaceous root hemiparasitic plant dish, which comprises the following steps:
(1) Collecting seeds with good maturity, sterilizing the surfaces of the seeds, and germinating the seeds in a light-temperature incubator to obtain robust aseptic seedlings;
(2) Cutting off main roots of the seedlings, and culturing in a light-temperature incubator to obtain root-cut seedlings;
(3) Selecting 2 root-cutting seedlings which emit new roots with uniform growth vigor again as a root-dividing material for establishing a root-dividing system in a vessel;
(4) Transferring the root-dividing material into a root-dividing device filled with a culture medium, and spreading 2 new roots on two sides of a partition plate to establish a root-dividing system;
(5) According to the purpose of the test, solid culture media with the same or different proportions can be added to the two sides of the root dividing device, or different biological combinations are arranged, so that the influence of different abiotic factors and biological factors on the growth and development of the root system can be inspected.
Further, the seeds with good maturity in the step (1) refer to the seeds with the shells changed into grey brown and full grains.
Further, the method for disinfecting the surface of the seeds in the step (1) comprises the following steps: placing the seeds in 70% alcohol for shock disinfection for 5 min, then washing with RO water until no alcohol smell exists, then shock disinfection for 10 min in 5wt% sodium hypochlorite, and flushing with sterile water in a super clean bench for more than 5 times until no disinfectant smell exists.
Further, the seedling in the step (2) is a seedling with cotyledon not breaking through seed coat and with radicle length of 0.5-1 cm.
Further, the root cutting position in the step (2) is at the root-stem junction, and the root cutting degree is to cut off all main roots.
Further, the length of the new root in the step (3) is 1-1.5 cm.
Further, no lateral root grows on the new root in the step (3).
Further, the root dividing device in the step (4) is a disposable sterile two-division plastic plate.
Further, the gelling agent used in the solid medium in the step (5) comprises agar, gellan gum or carrageenan.
Further, the non-biological factors in the step (5) include nitrogen, phosphorus, potassium, calcium, plant hormone auxin, gibberellin, cytokinin, abscisic acid, ethylene, strigolactone, sucker inducing compound DMBQ, sucrose, vanillin, vanillic acid, p-coumaric acid, p-hydroxybenzaldehyde, syringic acid, syringaldehyde, kaempferol and quercetin.
Further, the biological factors in the step (5) comprise host plants and microorganisms which have obvious influence on root hemiparasitic plants, wherein the host plants comprise the leguminous host plants of trifolium repens and alfalfa, the gramineous host plants of rice and brachypodium distachyon and the cruciferous host plants of arabidopsis thaliana, and the microorganisms comprise arbuscular mycorrhizal fungi and rhizobia.
It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the invention, as claimed.
The invention has the beneficial technical effects that:
the invention can evenly divide the root of the semi-parasitic plant seedling at the root of the herbaceous plant and reserve the capacity of the root system for generating the sucker, and a stable and reliable root dividing system is established in a short time. In the root dividing system built by the method, the root system shape and the occurrence level of the sucker can obviously respond to the nutrient supply change of a culture medium, the addition of sucker inducing substances or the change of biological environment, and the response trend is consistent with the performance of the whole root system without root division; the root system can clearly distinguish root behavior regulation and control modes which are difficult to distinguish in the whole root system.
Drawings
Fig. 1 shows the systematic regulation and control behavior of the root system of madder in Gansu province by using the internal division system of the vessel to help distinguish the form of the root system of madder in Gansu province by adding sucrose.
Detailed Description
Alternative embodiments of the present application will be described in more detail below with reference to the accompanying drawings. While the drawings show alternative embodiments of the present application, it should be understood that the present application may be embodied in various forms and should not be limited to the embodiments set forth herein. Rather, these embodiments are provided so that this disclosure will be thorough and complete, and will fully convey the scope of the disclosure to those skilled in the art.
The terminology used in the description of the invention herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used in this application and the appended claims, the singular forms "a", "an", and "the" are intended to include the plural forms as well, unless the context clearly indicates otherwise. It should also be understood that the term "and/or" as used herein refers to and encompasses any and all possible combinations of one or more of the associated listed items.
The method for establishing the root division system in the herbaceous root hemiparasitic plant dish in the invention is explained in detail with reference to the attached drawings, and concretely comprises the following steps:
the method for establishing the root dividing system in the herbaceous root hemiparasitic plant dish in the application of the invention comprises the following steps:
(1) Collecting seeds with good maturity, sterilizing the surfaces of the seeds, and germinating the seeds in a light-temperature incubator to obtain robust aseptic seedlings;
(2) Cutting off main roots of the seedlings, and culturing in a light-temperature incubator to obtain root-cut seedlings;
(3) Selecting 2 root-cutting seedlings which emit new roots with uniform growth vigor again as a root-dividing material for establishing a root-dividing system in a vessel;
(4) Transferring the root-dividing material into a root-dividing device filled with a culture medium, and spreading 2 new roots on two sides of a partition plate to establish a root-dividing system;
(5) According to the purpose of the test, solid culture media with the same or different proportions can be added to the two sides of the root dividing device, or different biological combinations are arranged, so that the influence of different abiotic factors and biological factors on the growth and development of the root system can be inspected.
Based on the understanding of the characteristics of herbaceous root semi-parasitic plants, the invention obtains strong seedlings by optimizing seeds, selects the seedling type, root cutting position and culture time suitable for root division, obtains the seedlings with uniform root division and capable of generating an aspirator, confirms the effectiveness and stability of the root division system in a vessel by comparing the root system morphological response of the root division system and a non-root division system (namely, the whole root), and solves the technical problem that the effective root division system of herbaceous root semi-parasitic plants is lacked at present.
In one embodiment of the present invention, the mature seeds in step (1) are full seeds with a shell changed into a grey brown color.
In one embodiment of the present application, the method for disinfecting the surface of the seed in step (1) comprises: placing the seeds in 70% alcohol for shock disinfection for 5 min, then washing with RO water until no alcohol smell exists, then shock disinfection for 10 min in 5wt% sodium hypochlorite, and flushing with sterile water in a super clean bench for more than 5 times until no disinfectant smell exists.
In one embodiment of the present application, the seedling in step (2) is a seedling whose cotyledon has not broken through the seed coat and whose radicle length is 0.5-1 cm.
In one embodiment of the present application, the root cutting position in the step (2) is at a root-stalk junction, and the degree of root cutting is to cut off all of the main roots.
In one embodiment of the present application, the new root length in step (3) is 1-1.5 cm.
In one embodiment of the present application, there is no lateral root outgrowth on the new root in step (3).
In one embodiment of the present application, the root-dividing device in step (4) is a disposable sterile two-compartment plastic plate.
In one embodiment of the present application, the gelling agent used in the solid medium described in step (5) comprises agar, gellan gum or carrageenan.
In one embodiment of the present application, the non-biological factors described in step (5) include the nutritional elements nitrogen, phosphorus, potassium, calcium, the plant hormones auxin, gibberellin, cytokinin, abscisic acid, ethylene, strigolactone, the haustorium inducing compounds DMBQ, sucrose, vanillin, vanillic acid, p-coumaric acid, p-hydroxybenzaldehyde, syringic acid, syringaldehyde, kaempferol and quercetin.
Further, the biological factors in the step (5) comprise host plants and microorganisms which have obvious influence on root hemiparasitic plants, wherein the host plants comprise the leguminous host plants of trifolium repens and alfalfa, the gramineous host plants of rice and brachypodium distachyon and the cruciferous host plants of arabidopsis thaliana, and the microorganisms comprise arbuscular mycorrhizal fungi and rhizobia.
For the sake of clarity, the following half-parasitic plant of root of Odonia tangutica of the genus Odonia of the family Orobanchaceae (Artemisia annua Gansu)Pedicularis kansuensis) The establishment of the root system and experimental verification are taken as examples, and the technical scheme in the embodiment of the invention is described in detail. It is noted that the details described herein are only some of the embodiments of the invention. All other embodiments obtained by anyone without creative work based on the embodiments of the present invention belong to the protection scope of the present invention.
Example 1 establishment of root division System in Gansu pedicularis Obertn seedling dish
The method for establishing the root dividing system of the herbaceous root hemiparasitic plant, namely the Gansu pedicularis, comprises the following steps:
(1) Preparing and germinating seeds: mature and full seeds of the mugwort in the middle of 2019 months in Bayin Bruker grassland of Xinjiang are collected, naturally dried at room temperature, and impurities are removed for later use. And (4) carrying out seed surface disinfection treatment before seed germination. Sterilizing seeds of herba pedicularis przewalskii in 70% alcohol by shaking for 5 min, washing with RO water until no alcohol smell, sterilizing in 5% sodium hypochlorite by shaking for 10 min, flushing in sterile water in superclean bench for more than 5 times, and placing the seeds on sterile filter paper. After the water on the surface of the seeds is dried, the seeds are inoculated on a water agar culture medium added with 0.8 wt percent one by using sterile tweezers, and 40 to 50 seeds are placed in each dish. Sealing the culture dish with seeds with 3M film, and placing in a 25/18 deg.C (day/night) light-temperature incubator for germination, wherein the light intensity is 22.2 μmol · m -2 ·s -1 12h light and 12h dark.
(2) And (3) culturing the root cutting seedlings: on the 9 th day after seed germination, selecting seedling with embryonic root length of 0.5-1 cm and cotyledon not break through seed coat, cutting off all main roots in super clean bench (cutting at root-stem junction), transferring the seedling to 0.8% water agar culture medium, and culturing in light-temperature incubator for 11 days.
(3) And (3) screening the root-divided seedlings: selecting seedlings which emit 2 new roots with uniform growth vigor again, have the length of the new roots of 1-1.5 cm and have no lateral roots on the new roots from the root cutting seedlings as root dividing materials, and establishing a dish root dividing system.
(4) Establishing an in-vessel root system: in a clean bench, the Gansu pedicel seedlings with the new root length of 1-1.5 cm (cultured for 11 days) are transferred into a disposable sterile two-division plastic plate filled with a culture medium, the seedlings are placed on a partition plate of the two-division plate, and 2 new roots are spread on two sides of the partition plate, so that a rooting system is established.
Comparative example 1-1 Effect of seedling type on root-dividing Effect of pedicularis mukoreana in Gansu
Comparative examples 1-1 were carried out strictly according to the conditions of example 1, except that the seedlings selected in comparative examples 1-1 were seedlings whose cotyledons did not break through the seed coat but whose radicle length was 1-1.5 cm or seedlings whose cotyledons broken through the seed coat and whose radicle length was 1-2 cm.
The comparative results show that the proportion of the homogeneously rooted material obtained in example 1 reached 36%, which is significantly higher than the seedlings in comparative example 1-1 (Table 1).
TABLE 1 Effect of seedling type on the rooting Effect of Gansu pedicularis
Comparative examples 1-2 Effect of root cutting position on root separating effect of Gansu pedicularis
Comparative examples 1-2 were carried out exactly as in example 1, except that the root cutting sites selected in comparative examples 1-2 were in the root elongation zone (about 0.5 cm below the root stalk junction) or the meristem zone (about 1 cm below the root stalk junction, near the root tip), respectively.
The comparison results show that the ratio of uniform rooting material obtained by cutting all the main roots at the root-stalk junctions in example 1 was 35%, which is significantly higher than the ratio of ideal rooting material obtained by cutting the roots in the root elongation zone or meristematic zone in comparative examples 1-2 (Table 2). In addition, the farther the root cutting position is from the root-stem joint, the more the number of the lateral roots subsequently produced by the seedling is, the more complex the root distribution is, and the worse the uniform root-dividing effect is. It follows that cutting all the main roots at the root-stalk junctions makes it easier to obtain seedlings with uniform root division.
TABLE 2 influence of root cutting position on root cutting effect of Gansu pedicularis
Comparative examples 1-3 Effect of cut root seedling culture time on root-dividing Effect of Gansu pedicularis
Comparative examples 1 to 3 were carried out exactly according to the conditions of example 1, except that the culture time after root cutting in comparative examples 1 to 3 was 7 days, 15 days and 20 days, respectively, to determine the culture time of the root-cutting seedling most suitable for the rooting operation.
The comparison results show that the proportion of ideally rooted seedlings decreases with the increase in the cut root seedling cultivation time (Table 3). Although the proportion of the uniform rooting seedlings is the highest on the 7 th day, the newly born root system is too short to be in contact with the culture medium for separating the two sides, the newly born root system is too long after 15 days, is easy to mutually wind and inconvenient for rooting operation, and the length of the newly born root system is about 1.5 cm when the seedling is cultured for 11 days, so that the seedling is in an ideal rooting state.
TABLE 3 influence of cut root seedling cultivation time on Gansu pedicel Effect of pedicel
In summary, when the root division system of the artemisia rupestris in Gansu is established, the seedlings with cotyledons not breaking through the seed coats and radicle length of 0.5-1 cm are preferably selected, all root cutting treatment is carried out at the root-stem junction, and the root cutting operation effect of the root cutting seedlings with new root length of about 1.5 cm (cultured for 11 days) is the best.
Example 2 root system differentiation of different modes of regulation and control of root morphology and haustoria of pedicularis margaritae in Gansu province by a root division system
To investigate whether the influence of sucrose on the growth and development of root system and generation of haustoria pedicularis in Gansu is local effect or systemic regulation and control effect, the method is based onThe method comprises the following steps of establishing a root dividing system of the artemisia rupestris in Gansu, and arranging different sucrose treatments on two sides of a partition plate of a two-grid plate, wherein the treatment steps are as follows: (1) adding 2% sucrose (+ S, + S) to both the left and right sides; (2) 2% sucrose was added on one side but not (+ S, -S) on the other side; (3) sucrose (-S, -S) is not added to the left and right sides. Each treatment was 30 dishes of 1 rooted seedling per dish. At the culture temperature of 25 ℃, the photoperiod of 12h light/12 h dark and the light intensity of 120 mu mol.m -2 ·s -1 The number of haustorium and the number of lateral roots are observed and recorded under a stereoscopic microscope after 14 days, and the total root length data is obtained by using WinRHIO analysis software after root system scanning. Three independent experiments were performed.
Comparative example 2 the whole root system could not distinguish the different modes of regulation and control of sucrose on root morphology and haustoria of pedicularis
Comparative example 2 was carried out exactly as in example 2, except that in comparative example 2, no rooting treatment was carried out, the entire root system was retained and the test was carried out in a plate of the same specification without partition.
The comparison results show that, in both the root division system of example 2 and the whole root system of comparative example 2, the number of haustorium, the number of lateral roots and the total root length of artemisia anomala in gansu were significantly increased after the addition of sucrose, compared to the treatment without the addition of sucrose (table 4), indicating that the addition of sucrose can effectively induce the occurrence of haustorium and lateral roots of artemisia anomala in gansu, and can also promote the increase of the total root length thereof. Although the number and the total root length of the haustorium and the lateral roots in the root system are less than those in the whole root system due to the removal of the main root, the haustorium can be normally generated by the root system in the root system, and the response rule of the root system to the addition of the cane sugar is consistent with that of the whole root system, which shows that the response behavior of the root system of the artemisia rupestris in Gansu can be accurately reflected by the experimental result carried out by the root system.
TABLE 4 response rule of suctor and root shape to sucrose in root system of pedicularis margaritae in Gansu province is consistent with that in whole root system
Note:* Shows the treatment with sucrose addition and the control without sucrose in the experimentP<There was a significant difference in the 0.05 level, indicating that the sucrose addition treatment compared to the no sucrose control in this group of experimentsP<There was a significant difference in the 0.01 level, indicating that the sucrose addition treatment compared to the no sucrose control in this group of experimentsP<There was a significant difference in the 0.001 level.
In the root system of example 2, when the treatment for separating both sides was the same, there was no significant difference in the root form index (table 5) and the occurrence level of the aspirator, indicating that the root system established by the method has uniform root division and stable and reliable results. In the whole root system of comparative example 2, only significant response of haustorium generation and root morphology to sucrose was observed, but it was not possible to judge whether the effect of sucrose on haustorium and root morphology was a local effect or a systemic effect. In the root system of example 2, it can be seen that the modes of sucroses on haustoria pedicularis in Gansu are different from the modes of root morphology regulation. The sucrose exerts an obvious induction effect in the aspect of haustorium generation and plays a role together through local induction and system regulation, because the addition of 2% of sucrose to one side of the root dividing system of the artemisia rupestris in Gansu can trigger the root system of the side (+ S side) to generate the haustorium, and can also induce the root system of the other side (-S side) without the addition of the sucrose to generate the haustorium, namely the sucrose can systematically induce the haustorium to generate through a long-distance signal path of root-stem-root; in the aspect of regulating and controlling root system morphology, the influence effect of sucrose shows different trends. When the local treatments on both sides of the root system are different (+ S, -S), the number of lateral roots and the total root length on both sides of the root system are very different. The side root number and the total root length on the + S side are similar to those in the (-S, -S) treatment, indicating that in the (+ S, -S) treatment, the + S side significantly suppresses the side root number and the total root length on the side, but greatly promotes the growth of the root system on the sucrose-free side. By utilizing the root system and assisting molecular detection and biochemical analysis means in the later period, deep research can be carried out on relevant mechanisms behind different regulation and control modes of the shapes of the suctors and the root systems by the sucrose.
TABLE 5 root division of Gansu pedicularis in different root rooms of the system haustorium generation and response of root morphology to sucrose treatment
Note: * The index on both sides of the experimental root of the group is shown inP<There was a significant difference in the 0.001 level.
Having described embodiments of the present application, the foregoing description is intended to be exemplary, not exhaustive, and not limited to the embodiments disclosed. Many modifications and variations will be apparent to those of ordinary skill in the art without departing from the scope and spirit of the described embodiments. The terminology used herein is chosen in order to best explain the principles of the embodiments, the practical application, or improvements made to the technology in the marketplace, or to enable others of ordinary skill in the art to understand the embodiments disclosed herein.
Claims (7)
1. A method for establishing a root division system in a herbaceous root hemiparasitic plant dish is characterized by comprising the following steps:
(1) Collecting seeds with good maturity, sterilizing the surfaces of the seeds, and germinating the seeds in a light-temperature incubator to obtain robust sterile seedlings;
(2) Cutting off all main roots of the seedlings along the root-stem combination part, and culturing in a light-temperature culture box for 11 days to obtain root-cut seedlings, wherein the seedlings have cotyledon not break through seed coat and have radicle length of 0.5-1 cm;
(3) Selecting 2 re-rooted seedlings which have uniform new roots and have no lateral roots to grow as a root-dividing material for establishing a root-dividing system in a vessel, wherein the length of the new roots is 1-1.5 cm;
(4) Transferring the root-dividing material into a root-dividing device filled with a culture medium, spreading 2 new roots on two sides of a partition plate, and establishing a root-dividing system;
(5) According to the purpose of the test, solid culture media with the same or different proportions can be added to the two sides of the root dividing device, or different biological combinations are arranged, so that the influence of different abiotic factors and biological factors on the growth and development of the root system can be inspected.
2. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the seeds with good maturity in the step (1) are seeds with grayish brown shells and full grains.
3. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the method for disinfecting the surface of the seeds in the step (1) comprises the following steps: the seeds are placed in 70% alcohol for shake disinfection for 5 min, then washed by RO water until no alcohol smell exists, shaken in 5wt% sodium hypochlorite for disinfection for 10 min, and washed by sterile water in a super clean bench for more than 5 times until no disinfectant smell exists.
4. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the root dividing device in the step (4) is a disposable sterile two-division plastic plate.
5. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the gel used in the solid culture medium in the step (5) comprises agar, gellan gum or carrageenan.
6. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the non-biological factors in the step (5) comprise nutrient elements, plant hormones and haustorium inducing compounds, wherein the nutrient elements comprise nitrogen, phosphorus, potassium and calcium, the plant hormones comprise auxin, gibberellin, cytokinin, abscisic acid, ethylene and strigolactone, and the haustorium inducing compounds comprise DMBQ, sucrose, vanillin, vanillic acid, p-coumaric acid, p-hydroxybenzaldehyde, syringic acid, syringaldehyde, kaempferol and quercetin.
7. The method of establishing a herbaceous root hemiparasitic plant in-dish root system as claimed in claim 1, wherein: the biological factors in the step (5) comprise host plants and microorganisms which have obvious influence on root hemiparasitic plants, wherein the host plants comprise leguminous host plants such as trifolium repens and alfalfa, gramineous host plants such as rice and brachypodium distachyon and cruciferae host plants such as arabidopsis thaliana, and the microorganisms comprise arbuscular mycorrhizal fungi and rhizobia.
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