CN115968762A - Novel variety cultivation method for breaking self-incompatibility of kochia scoparia - Google Patents

Abstract

The invention provides a new species cultivation method for breaking self-incompatibility of the rubberella through a root segment asexual rapid propagation technology, which achieves the purpose of rapidly and asexually propagating a plurality of plants from a single plant by controlling factors such as moisture, temperature, exogenous hormone and the like in the environment, and has the advantages of simple method, low requirement on experimental environment, rapid seedling formation and more than 80 percent of seedling rate. Meanwhile, a plurality of rapidly-bred plants are isolated for free pollination, so that free mating of different plants with the same genotype is realized, the problem of breaking self-incompatibility of the kochia scoparia can be solved, a new realization path and a new realization method are established for purification and new variety breeding of kochia scoparia strains, and the method has great potential development value for large-scale planting of the kochia scoparia.

Description

Novel variety cultivation method for breaking self-incompatibility of kochia scoparia

Technical Field

The invention belongs to the technical field of biology, particularly relates to the technical field of rubber grass breeding, and more particularly relates to a new variety breeding method for breaking self-incompatibility of rubber grass by root segment asexual rapid propagation technology and the technical field of application.

Technical Field

Natural rubber, steel, coal and petroleum are called four basic raw materials, and are very important strategic resources. At present, the consumption of natural rubber in China is about 600 ten thousand tons, and the natural rubber accounts for over 40 percent of the whole world. Wherein, the domestic yield accounts for only 13 percent of the consumption, and the external dependence is more than 80 percent. The natural rubber is mainly produced from hevea trees planted in tropical rainforest regions suitable for damp and hot weather, but the regions suitable for the growth of the hevea trees in China are very limited, so the natural rubber is an important agricultural product for restricting the economic development and strategic safety of China. Hedychium notatum (Taraxacumkoksaghyz Rodin, TKS, also known as Taraxacum russiamensis or Taraxacum kok. Var. Mongolicum (Maxim.) DC.) C.of Taraxacum of Compositae) is a perennial herb. The rubber is originally produced in the places of Hassakestan, gill Ji Sisi, russia, xinjiang and the like, and becomes a Hevea rubber substitute crop with great development prospect due to the advantages of high natural rubber content of roots, excellent quality, strong adaptability and the like. The rubber grass has the characteristics of wide adaptability, short rubber production period, easy mechanical operation, effective utilization of saline-alkali soil and the like, and is an advantageous crop which can solve the problem of long-term self-sufficiency of natural rubber in China and has potential development value.

The chromosome number of the Hevea brasiliensis is 2n =16, the Hevea brasiliensis has complete flowers capable of forming normal female and male gametes, and the Hevea brasiliensis cannot fruit completely during self-pollination, so that the variety breeding of the Hevea brasiliensis is greatly influenced.

Disclosure of Invention

Aiming at the problem that the prior art can not fruit completely when the rubber grass is self-pollinated and causes great influence on variety breeding of the rubber grass, the invention controls factors such as moisture, temperature, exogenous hormone and the like in the environment, achieves the purpose that the rubber grass is rapidly propagated by a single plant to obtain a plurality of plants by asexual propagation, realizes the self mating of different plants with the same genotype, and solves the problem of breaking the self incompatibility of the rubber grass. Has great potential development value for the scale planting of the rubber grass.

In order to realize the purpose of the invention, the technical scheme of the invention is as follows:

the application provides a new variety cultivation method for breaking self-incompatibility of kochia scoparia, which is obtained by adopting the following steps:

(1) Taking a clean culture dish with the diameter of about 150mm, and putting 1 piece of dry filter paper with the same specification on the culture dish; selecting excellent individual plants of the kochia scoparia which meet the breeding target, cleaning soil at the root part of the kochia scoparia with clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the root part of the kochia scoparia with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the upper end and the lower end of the root part in a culture dish according to morphology, treating the root part by adopting phytohormone, and standing the root part overnight;

(2) Adding clear water into a special vegetable seedling culture medium which is sold in the market and mainly comprises grass carbon, uniformly stirring, kneading the medium into a ball, falling onto the ground, naturally dispersing, and then filling the mixed medium into a plastic flowerpot or a seedling culture hole tray with 72 holes; placing the flowerpot or seedling raising tray filled with the matrix into a tray of about 5cm; taking out the overnight rubber grass root sections treated in the step (1), inserting the lower ends of the overnight rubber grass root sections into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm;

(3) Filling water into the tray in the step (2) after the root segments of the rubber grasses are cut, and enabling the substrate to absorb water from the tray to keep the proper humidity of the substrate so as to facilitate the rooting of the rubber grasses; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced;

(4) Observing the germinated rubber grass tissues treated in the step (3) after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray;

(5) Observing that the root segment of the rubberella tabacum in the step (4) generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubberella tabacum in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubberberis are prevented from being infected by mould, rotted and dead due to overhigh humidity;

(6) Transplanting the rapidly-propagated rubber grass in the step (5) to a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting.

(7) Fully wrapping the root systems of the rubberweeds cultured in the step (6) by seedling culture substrates after 45 days, taking out the rubberweeds together with the seedling culture substrates, transplanting the rubberweeds into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberweeds after 7 days, and removing the sunshade net;

(8) When the rubber grass treated in the step (6) buds, using an insect-proof net to make a small arched shed for isolation, preventing powder from being mixed among different materials of the rubber grass, wherein 10-12 points are arranged in the morning each day, the flowers of the rubber grass are completely opened, the flowers are artificially pollinated in an auxiliary way when the stigma is in a Y shape, and seeds can be harvested about 12 days after blooming;

(9) And (3) selecting materials by the method in the steps (1) to (8), carrying out rapid asexual propagation for multiple generations, finally realizing the purification of the rubber grass materials, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the rubber grass with high quality.

Furthermore, the plant hormone can adopt gibberellin, indoleacetic acid and indolebutyric acid.

Furthermore, the phytohormone can be gibberellin, indoleacetic acid and indolebutyric acid respectively, and the concentration is 80-240mg/L.

Preferably, the phytohormone can be gibberellin, indoleacetic acid and indolebutyric acid respectively, and the concentration is 160mg/L,

Preferably, 16-240mg/L gibberellin and 80-240mg/L indolebutyric acid are adopted as the phytohormone.

More preferably, 160mg/L gibberellin and 240mg/L indolebutyric acid are used as the phytohormones.

Furthermore, the application also provides the application of the new variety cultivation method for breaking the self-incompatibility of the kochia scoparia in the new variety cultivation of the kochia scoparia.

By adopting the technical scheme provided by the invention, the invention has the following beneficial effects

(1) The invention provides a new variety cultivation method for breaking self-incompatibility of the hevea brasiliensis by a root segment asexual rapid propagation technology, which achieves the purpose of rapidly and asexually propagating a plurality of plants from a single plant by controlling factors such as moisture, temperature, exogenous hormone and the like in the environment.

(2) By applying the new variety cultivation method for breaking the self-incompatibility of the rubbergrass through the root segment asexual rapid propagation technology, the operation method is simple, the requirement on the experimental environment is low, the seedling is fast, and the seedling rate reaches more than 80%. Meanwhile, a plurality of rapidly propagated plants are isolated and pollinated freely, so that different plants with the same genotype are mated freely, the problem of breaking the self-incompatibility of the kochia scoparia is solved, a new realization path and method are established for purification of kochia scoparia strains and breeding of new varieties, and the method has great potential development value for large-scale planting of the kochia scoparia.

Drawings

FIG. 1 is a graph showing the effect of different concentrations of gibberellin on the germination and rooting rate of root segments at different parts.

FIG. 2 is a graph showing the effect of different concentrations of indoleacetic acid on the germination and rooting rate of root segments at different positions.

FIG. 3 is a graph showing the influence of different concentrations of indolebutyric acid on the germination and rooting rate of root segments at different positions.

FIG. 4 is a graph showing the results of the control experiment.

FIG. 5 is a graph showing the effect of different GA3+ IBA concentrations on the germination and rooting rates of root segments at different positions.

FIG. 6 shows examples of asexual rapid propagation of root segments. Wherein panel A shows individual plant selection for RUBENCAO; FIG. B shows the cut-off of the grass roots placed in a petri dish with filter paper; FIG. C shows that the primordium of the leaf bud is just appeared after about 7 days of cutting culture; FIG. D shows the yellow nearly transparent new leaf at around 13 days old; panel E shows the leaves turning green for about 16 days to form new leaves of RUBENCAO.

FIG. 7 is a field diagram of the propagation of the root segment of kochia kochiana.

FIG. 8 shows isolated pollination of a rubber grass net chamber.

FIG. 9 is a graph showing the flowering habit observation and pollination of Hevea brasiliensis.

The specific implementation mode is as follows:

the following examples are intended to further illustrate the present invention but are not to be construed as limiting the invention. It is within the scope of the present invention to modify or replace methods, steps or conditions of the present invention without departing from the spirit and substance of the present invention.

The experimental procedures used in the following examples are all conventional procedures unless otherwise specified. Materials, reagents and the like used in the following examples are commercially available unless otherwise specified.

The first embodiment is as follows: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

The method for cultivating a new variety of the rubberella multocida for breaking self-incompatibility of the rubberella multocida is obtained by the following steps:

(1) And taking a clean culture dish with the diameter of about 150mm, and putting 1 piece of dry filter paper with the same specification on the culture dish. Selecting excellent individual plants of the kochia scoparia which meet the breeding target, cleaning soil at the root part of the kochia scoparia with clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the root part of the kochia scoparia with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the upper end and the lower end of the root part in a culture dish according to morphology, treating the root part by adopting phytohormone, and standing the root part overnight;

(2) Adding clear water into a special vegetable seedling culture medium which is sold in the market and mainly contains turf, uniformly stirring the clear water and the special vegetable seedling culture medium to grab the medium, kneading the medium into a cluster, dropping the cluster onto the ground and naturally dispersing the cluster, and then filling the mixed medium into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the overnight rubber grass root sections treated in the step (1), inserting the lower ends of the overnight rubber grass root sections into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm;

(3) Filling water into the tray in the step (2) after the root segments of the rubber grasses are cut, and enabling the substrate to absorb water from the tray to keep the proper humidity of the substrate so as to facilitate the rooting of the rubber grasses; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced;

(4) Observing the germinated rubber grass tissues treated in the step (3) after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray;

(5) Observing that the root segment of the rubberella tabacum in the step (4) generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubberella tabacum in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubberberis are prevented from being infected by mould, rotted and dead due to overhigh humidity;

(6) Transplanting the rapidly-propagated rubber grass in the step (5) to a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting.

(7) Fully wrapping seedling raising substrates with root systems of the rubber grasses cultured in the step (6) after about 45 days, taking out the rubber grasses together with the seedling raising substrates, transplanting the rubber grasses into a test cell, covering a sunshade net, watering thoroughly, reviving the rubber grasses after about 7 days, and removing the sunshade net;

(8) When the rubber grass treated in the step (6) buds, using an insect-proof net to make a small arched shed for isolation, preventing powder from being mixed among different materials of the rubber grass, wherein 10-12 points are arranged in the morning each day, the flowers of the rubber grass are completely opened, the flowers are artificially pollinated in an auxiliary way when the stigma is in a Y shape, and seeds can be harvested about 12 days after blooming;

(9) And (3) selecting materials by the method in the steps (1) to (8), carrying out rapid asexual propagation for multiple generations, finally realizing the purification of the rubber grass materials, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the rubber grass with high quality.

The second embodiment: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the rubber grass which meet the breeding target, cleaning soil at the root part of the rubber grass by using clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the rubber grass root with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the root sections into a culture dish according to the morphological upper end and the lower end, treating by adopting 80mg/L gibberellin or indoleacetic acid or indolebutyric acid, and standing overnight; adding clear water into a special substrate (main component: turf) for vegetable seedling culture sold in the market, uniformly stirring, picking up the substrate, kneading the substrate into a cluster, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; the tray after the root segments of the rubber grasses are cut is filled with water, and the substrate can absorb water from the tray to keep the proper humidity of the substrate, so that the rubber grasses can take root conveniently; after the flowerpot or the seedling culture hole tray is fully covered by a commercially available transparent film after being filled with water, a plurality of holes are uniformly drilled on the film to facilitate ventilation, so that the condition that the upper growing point of the root section cannot be dehydrated too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated rubber grass after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray; observing that the root segment of the rubber grass generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubber grass in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubberberis are prevented from being infected by mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass to a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubber grass buds, the small arched shed is made of an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubber grass, 10-12 points are arranged in the morning, the rubber grass flowers are completely opened, the stigma is in a Y shape, the flowers are pollinated in an auxiliary mode, and seeds can be collected after about 12 days after the flowers bloom; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia.

Example three: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the kochia scoparia which meet the breeding target, cleaning soil at the root part of the kochia scoparia with clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the root part of the kochia scoparia with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the upper end and the lower end in a culture dish according to morphology, treating the root parts by adopting 160mg/L gibberellin or indoleacetic acid or indolebutyric acid, and standing the root parts overnight; adding clear water into a special substrate (main component: turf) for vegetable seedling culture sold in the market, uniformly stirring, picking up the substrate, kneading the substrate into a cluster, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; filling water into the tray after the cuttage of the rubber grass root segments is finished, and enabling the substrate to absorb water from the tray so as to keep the proper humidity of the substrate, so that the rubber grass root segments can take root conveniently; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated rubber grass after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray; observing that the root segment of the rubber grass generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubber grass in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray still needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubber grass are prevented from being infected with mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass into a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubberella appears, a small arched shed is made by using an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubberella, 10-12 points are arranged in the morning, the flowers of the rubberella are completely opened, the flower is pollinated by manual pollination when the stigma is in a Y shape, and seeds can be collected about 12 days after the flowers are bloomed; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia.

Example four: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the kochia scoparia which meet the breeding target, cleaning soil at the root part of the kochia scoparia with clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the root part of the kochia scoparia with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the upper end and the lower end in a culture dish according to morphology, treating the root parts by adopting 240mg/L gibberellin or indoleacetic acid or indolebutyric acid, and standing the root parts overnight; adding clear water into a special substrate (main component: turf) for vegetable seedling culture sold in the market, uniformly stirring, picking up the substrate, kneading the substrate into a cluster, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; the tray after the root segments of the rubber grasses are cut is filled with water, and the substrate can absorb water from the tray to keep the proper humidity of the substrate, so that the rubber grasses can take root conveniently; after the flowerpot or the seedling culture hole tray is fully covered by a commercially available transparent film after being filled with water, a plurality of holes are uniformly drilled on the film to facilitate ventilation, so that the condition that the upper growing point of the root section cannot be dehydrated too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated rubber grass after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray; observing that leaf bud primordium can be differentiated in about 7 days generally, yellow nearly transparent new leaves can grow in about 13 days, and leaves can turn green to form new leaves of the Hevea brasiliensis in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubberberis are prevented from being infected by mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass into a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubberella appears, a small arched shed is made by using an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubberella, 10-12 points are arranged in the morning, the flowers of the rubberella are completely opened, the flower is pollinated by manual pollination when the stigma is in a Y shape, and seeds can be collected about 12 days after the flowers are bloomed; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia.

Example five: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the kochia scoparia which meet the breeding target, cleaning soil at the root part of the kochia scoparia with clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the root part of the kochia scoparia with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the upper end and the lower end in a culture dish according to morphology, treating the root parts by adopting 160mg/L gibberellin and 80mg/L indolebutyric acid, and standing the root parts overnight; adding clear water into a special substrate (main component: turf) for vegetable seedling culture sold in the market, uniformly stirring, picking up the substrate, kneading the substrate into a cluster, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; filling water into the tray after the cuttage of the rubber grass root segments is finished, and enabling the substrate to absorb water from the tray so as to keep the proper humidity of the substrate, so that the rubber grass root segments can take root conveniently; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated tarragon, and properly irrigating before the germination of the tarragon according to the evaporation condition of water in the tray; observing that leaf bud primordium can be differentiated in about 7 days generally, yellow nearly transparent new leaves can grow in about 13 days, and leaves can turn green to form new leaves of the Hevea brasiliensis in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray still needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubber grass are prevented from being infected with mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass into a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubberella appears, a small arched shed is made by using an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubberella, 10-12 points are arranged in the morning, the flowers of the rubberella are completely opened, the flower is pollinated by manual pollination when the stigma is in a Y shape, and seeds can be collected about 12 days after the flowers are bloomed; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia.

Example six: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the rubber grass which meet the breeding target, cleaning soil at the root part of the rubber grass by using clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the rubber grass root with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the root sections into a culture dish according to the morphological upper end and the lower end, treating by using 160mg/L gibberellin and 160mg/L indolebutyric acid, and standing overnight; adding clear water into a special substrate (main component: grass carbon) for vegetable seedling culture sold in the market, uniformly stirring, kneading the substrate into a ball, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; filling water into the tray after the cuttage of the rubber grass root segments is finished, and enabling the substrate to absorb water from the tray so as to keep the proper humidity of the substrate, so that the rubber grass root segments can take root conveniently; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated rubber grass after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray; observing that the root segment of the rubber grass generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubber grass in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can continuously grow on the roots, and new rubber grass plants gradually grow; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubberberis are prevented from being infected by mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass into a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30%, the temperature is 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening off the seedling and waits for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubberella appears, a small arched shed is made by using an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubberella, 10-12 points are arranged in the morning, the flowers of the rubberella are completely opened, the flower is pollinated by manual pollination when the stigma is in a Y shape, and seeds can be collected about 12 days after the flowers are bloomed; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia.

Example seven: novel variety cultivation method for breaking self-incompatibility of kochia scoparia

A clean culture dish with the diameter of about 150mm is taken, and 1 piece of dry filter paper with the same specification is placed on the culture dish. Selecting excellent individual plants of the rubber grass which meet the breeding target, cleaning soil at the root part of the rubber grass by using clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the rubber grass root with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the root sections into a culture dish according to the morphological upper end and the lower end, treating by using 160mg/L gibberellin and 240mg/L indolebutyric acid, and standing overnight; adding clear water into a special substrate (main component: turf) for vegetable seedling culture sold in the market, uniformly stirring, picking up the substrate, kneading the substrate into a cluster, falling onto the ground, naturally dispersing, and then filling the mixed substrate into a plastic flowerpot or a seedling culture hole tray with 72 holes. The flowerpot or seedling raising tray filled with the matrix is put into a tray of about 5 cm. Taking out the treated overnight rubber grass root segments, inserting the lower ends of the rubber grass root segments into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm; filling water into the tray after the cuttage of the rubber grass root segments is finished, and enabling the substrate to absorb water from the tray so as to keep the proper humidity of the substrate, so that the rubber grass root segments can take root conveniently; after the flowerpot or the seedling culture hole tray is fully covered by a commercially available transparent film after being filled with water, a plurality of holes are uniformly drilled on the film to facilitate ventilation, so that the condition that the upper growing point of the root section cannot be dehydrated too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced; observing the tissue of the germinated rubber grass after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray; observing that the root segment of the rubber grass generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubber grass in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray still needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubber grass are prevented from being infected with mould, rotted and dead due to overhigh humidity; transplanting the rapidly-propagated rubber grass to a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting; fully wrapping seedling-raising substrates with roots of the rubberella tabacum after being cultured for about 45 days, taking out the rubberella tabacum seedlings together with the seedling-raising substrates, transplanting the rubberella tabacum seedlings into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberella tabacum seedlings for about 7 days, and removing the sunshade net; when the rubberella appears, a small arched shed is made by using an insect-proof net for isolation, powder is prevented from being mixed among different materials of the rubberella, 10-12 points are arranged in the morning, the flowers of the rubberella are completely opened, the flower is pollinated by manual pollination when the stigma is in a Y shape, and seeds can be collected about 12 days after the flowers are bloomed; the method is used for selecting materials, carrying out multi-generation asexual rapid propagation, finally realizing the purification of the kochia scoparia material, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the high-quality kochia scoparia. The details of the growth and experimental conditions are shown in the accompanying figures 6, 7, 8 and 9.

Example seven: comparison of the effects of different phytohormones

Based on the above examples one to six, the effects of different phytohormone treatments were compared.

1 materials and methods

1.1 test materials

The rubber grass root section is made of a Torontis variety, and the sampling site is the Yili area of Xinjiang. The filter paper used was a normal filter paper, and the diameter of the glass petri dish was 100mm. The temperature of the artificial climate chamber is 28 ℃ and the relative humidity is 60 percent.

1.2 test methods

The test site is in the first floor of cotton building of academy of agricultural sciences in Xinjiang, the roots of kochia scoparia are divided into three parts, namely an upper part, a middle part and a lower part, 1.5-2 cm long root sections are cut, a filter paper germinating bed is soaked, 20 kochia scoparia root sections are placed in each culture dish for rooting and germinating tests, and each concentration is divided into an upper part, a middle part and a lower part which are placed in three culture dishes. The single hormone culture dishes are respectively added with 1ml of 80mg/L, 160mg/L and 240mg/L of GA3, IAA and IBA 3 hormones with concentration gradient and 4ml of distilled water. 0.5ml of GA380mg/L and 160mg/L was mixed with 0.5ml of IBA and IAA 80mg/L, 160mg/L and 240mg/L in a dish containing mixed hormones, and 4ml of distilled water was added to each dish. Then placing the mixture into a phytotron with the temperature of 28 +/-0.5 ℃ and the relative humidity of 60 percent to observe the germination and rooting conditions. Supplementing 1ml of distilled water every day, taking the standard that the root segments grow new roots more than 2mm as rooting standards, taking the standard that the root segments grow new leaves and show green as germination standards, and counting the number of the root segments for rooting and germination day by day until the number of the root segments for rooting and germination is unchanged.

1.3 data statistics and processing

The calculation formula is as follows:

germination percentage (%) = total number of germinating root segments/total number of test root segments × 100%;

rooting rate (%) = total number of rooted root segments/total number of test root segments × 100%.

2 results and analysis

2.1 Effect of Single hormone on sprouting and rooting of root segments at different positions

The root segments of the kojiki are soaked by 3 hormones of gibberellin, indoleacetic acid and indolebutyric acid in different concentrations, and the rooting and sprouting conditions of the root segments of the kojiki are observed and recorded by taking distilled water as a control. The test result shows that: the rubber grass root segment has certain totipotency, and can differentiate to grow new roots and buds under the soaking treatment of hormones and distilled water to grow into a complete plant. Referring to FIG. 1, the germination potential and germination rate of the root segment of the kochia scoparia at a concentration of 160mg/L were significantly higher than other concentrations under gibberellin treatment; under the treatment of different concentrations, the root segments of the rubber grass partially take root, and under the treatment of different concentrations without obvious rules, the root germination rate of the lower root segments of the rubber grass close to the root crown end is obviously higher than that of the upper and middle root segments, the root germination rate of the lower root segments treated by 160mg/L gibberellin reaches the highest, the germination rate is 65 percent, and the root germination rate is 20 percent. Referring to the attached figure 2, under the treatment of indoleacetic acid, the germination rate of the root segment of the rubber grass is obviously lower than that of gibberellin, the highest germination rate of the middle root segment of the rubber grass under the treatment of 160mg/L concentration reaches 35 percent, the rooting aspect is obviously better than that of the middle root segment of the rubber grass under the treatment of gibberellin, and the highest rooting rate of the middle root segment of the rubber grass under the treatment of 240mg/L concentration reaches 50 percent; the rooting rate of the middle and lower root segments is obviously higher than that of the upper root segment, and the germination rates of the root segments at different parts have no obvious rule; the rooting rate and the germination rate have no obvious difference. Referring to the attached figure 3, under the treatment of indolebutyric acid, the germination rates of the rubber grass root segments are obviously reduced through the treatment of different concentrations, the germination rate of the rubber grass root segments of the great part is reduced to 0, and the rooting rate is obviously increased to 100 percent at most; wherein, the rooting performance of the cord grass root segments treated by the concentration of 160mg/L is stable, the rooting rate of the upper, middle and lower root segments reaches more than 50 percent, and the rooting rate of the lower root segment is the highest. Referring to the attached figure 4, the rooting and germination rates of the lower root segment are obviously higher than those of the middle and upper root segments under the treatment of the control distilled water, and the rooting rate and the germination rate have no obvious difference. In a whole view, the rooting and sprouting of the rubber grass root segment does not show obvious promotion and inhibition effects under the treatment of the indoleacetic acid and the control distilled water, gibberellin can promote the sprouting of the rubber grass root segment, and the indolebutyric acid can promote the rooting of the rubber grass root segment.

2.1.2 Effect of Mixed hormones on the sprouting and rooting of root segments at different positions

The analysis result of 2.1.1 shows that different hormones have different degrees of inhibition and promotion effects on the rooting and sprouting of the rubber grass root segment. Therefore, in the process of culturing the rubber grass root segment, two hormones with different concentrations are added for treatment at the same time, so as to obtain the optimal hormone concentration ratio for the rooting and sprouting of the rubber grass root segment. Referring to figure 5, the root segments treated by the mixed hormones GA3+ IAA and GA3+ IBA can emit new roots and buds, but the obvious difference exists, and the test treatment of failing to root and sprout is not carried out. The upper, middle and lower parts treated by GA3+ IBA (160 mg/L +80 mg/L), GA3+ IBA (160 mg/L +240 mg/L), GA3+ IAA (80 mg/L +80 mg/L) and GA (80 mg/L +160 mg/L) all have root segments to completely grow new roots and buds. Wherein, the root section of GA3+ IBA (160 mg/L +240 mg/L) has more stable rooting and sprouting, and the rooting and sprouting rate is higher and is obviously higher than that of the control distilled water treatment.

As described above, the present invention can be preferably implemented, and the above-mentioned embodiments are merely illustrative of the preferred embodiments of the present invention, and do not limit the scope of the present invention, and various changes and modifications made to the technical solution of the present invention by those skilled in the art without departing from the spirit of the present invention should fall within the protection scope defined by the present invention.

Claims (7)

1. A new variety cultivation method for breaking self-incompatibility of Hevea brasiliensis is characterized in that the cultivation method comprises the following steps:

(1) Taking a clean culture dish with the diameter of about 150mm, and putting 1 piece of dry filter paper with the same specification on the culture dish; selecting excellent individual plants of the rubber grass which meet the breeding target, cleaning soil at the root part of the rubber grass by using clear water, removing leaves at the overground part, completely stripping the outer layer tissues of the rotten root part, cutting the rubber grass root with the diameter of more than 1mm into root sections of 1-2cm by using a sharp blade, neatly placing the root sections in a culture dish according to the morphological upper end and the lower end, treating by using phytohormone, and standing overnight;

(2) Adding clear water into a special vegetable seedling culture medium which is sold in the market and mainly contains turf, uniformly stirring the clear water and the special vegetable seedling culture medium to grab the medium, kneading the medium into a cluster, dropping the cluster onto the ground and naturally dispersing the cluster, and then filling the mixed medium into a plastic flowerpot or a seedling culture plug tray with 72 holes; putting the flowerpot or seedling raising tray filled with the matrix into a tray of about 5cm; taking out the overnight rubber grass root sections treated in the step (1), inserting the lower ends of the overnight rubber grass root sections into a seedling culture substrate according to morphology, and exposing the substrate at the upper ends by about 0.5cm;

(3) Filling water into the tray in the step (2) after the root segments of the rubber grasses are cut, and enabling the substrate to absorb water from the tray to keep the proper humidity of the substrate so as to facilitate the rooting of the rubber grasses; after the flower pot or the seedling culture hole tray is fully covered by a commercially available transparent film after the water is filled, a plurality of holes are uniformly drilled on the film to facilitate ventilation, and the condition that the upper growing point of the root section cannot lose water too quickly to influence germination can be effectively ensured; under the condition of ensuring the humidity, the indoor temperature is controlled to be 25-28 ℃, and the too fast evaporation of water caused by direct solar radiation is reduced;

(4) Observing the germinated rubber grass tissues treated in the step (3) after cuttage, and properly irrigating water before germination of the rubber grass according to the evaporation condition of water in the tray;

(5) Observing that the root segment of the rubberella tabacum in the step (4) generally can differentiate into leaf bud primordium in about 7 days, yellow and nearly transparent new leaves grow in about 13 days, and the leaves turn green to form new leaves of the rubberella tabacum in about 16 days; generally more than 80% of root segments can grow new leaves in the time, new root hairs can be grown at the roots successively, and new rubber grass plants are grown gradually; when yellow and nearly transparent new leaves grow, irrigation needs to be reduced, if a large amount of water in the tray still needs to be poured out in time, the humidity in the substrate is reduced, and the new plants of the rubber grass are prevented from being infected with mould, rotted and dead due to overhigh humidity;

(6) Transplanting the rapidly-propagated rubber grass in the step (5) to a test field for about 45 days; the root system of the rubber grass plant which is about 16 days is not completely developed, the soil humidity is controlled to be about 30 percent, the temperature is controlled to be 25-28 ℃, the plant continues to grow for about 20 days, and then the plant is transferred to the outdoor for hardening seedlings to wait for transplanting;

(7) Fully wrapping the root systems of the rubberweeds cultured in the step (6) by seedling culture substrates after 45 days, taking out the rubberweeds together with the seedling culture substrates, transplanting the rubberweeds into a test cell, covering a sunshade net, watering thoroughly, reviving the rubberweeds after 7 days, and removing the sunshade net;

(8) When the rubber grass treated in the step (6) buds, using an insect-proof net to make a small arched shed for isolation, preventing powder from being mixed among different materials of the rubber grass, wherein 10-12 points are arranged in the morning each day, the flowers of the rubber grass are completely opened, the flowers are artificially pollinated in an auxiliary way when the stigma is in a Y shape, and seeds can be harvested about 12 days after blooming;

(9) And (3) selecting materials by the method in the steps (1) to (8), carrying out rapid asexual propagation for multiple generations, finally realizing the purification of the rubber grass materials, and mixing inbred lines with similar agronomic characters and higher glue content for free mating to obtain a new variety of the rubber grass with high quality.

2. The method of claim 1, wherein said plant hormone is selected from the group consisting of gibberellin, indoleacetic acid, indolebutyric acid.

3. The method for breeding a new variety of plants to break the self-incompatibility of Hevea brasiliensis as claimed in claim 2, wherein said plant hormones can be gibberellin, indoleacetic acid, indolebutyric acid, respectively, at a concentration of 80-240mg/L.

4. The method as claimed in claim 3, wherein the plant hormones are selected from gibberellin, indoleacetic acid and indolebutyric acid at a concentration of 160mg/L.

5. The method as claimed in claim 2, wherein the plant hormone is selected from the group consisting of 160-240mg/L gibberellin and 80-240mg/L indolebutyric acid.

6. The method of claim 2, wherein 160mg/L gibberellin and 240mg/L indolebutyric acid are used as the plant hormones.

7. Use of the method of any one of claims 1 to 5 for breeding a new variety of kombucha that breaks self-incompatibility.

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