阅读说明：本技术 一种复苏植物垫状卷柏的珠芽繁殖方法 (Pearl bud propagation method for reviving selaginella pulvinata ) 是由 余蓉培 杨春梅 李绅崇 吴丽芳 阮继伟 单芹丽 吴旻 于 2019-12-16 设计创作，主要内容包括：一种复苏植物垫状卷柏的珠芽繁殖方法,包括：垫状卷柏珠芽的诱导培养、珠芽的穴盘苗培养、珠芽穴盘苗的定植以及定植后的栽培管理。本发明成功以野生垫状卷柏2～3年生植株为材料,通过对栽培基质、光照条件以及水分条件的控制,经过5～6个月的珠芽诱导培养,珠芽诱导率达100％,单个成熟枝条的平均珠芽形成数在20个珠芽以上。定植后的垫状卷柏珠芽在弱光照温室、强光照温室以及露天场地等不同栽培环境中进行分步驯化培养,垫状卷柏珠芽的定植成活率可达93％以上,填补了垫状卷柏人工繁殖技术的空白,对复苏植物垫状卷柏的野生种群保护和资源开发利用具有重要意义。(A method for reproducing bulbil of herba Selaginellae Doederleinii comprises: the method comprises the following steps of induced culture of selaginella pulvinata bulbels, plug seedling culture of the bulbels, field planting of the plug seedlings of the bulbels and cultivation management after the field planting. According to the method, 2-3 years old plants of wild selaginella pulvinata maxim are successfully used as materials, the culture medium, the illumination condition and the moisture condition are controlled, the pearl bud induction culture is carried out for 5-6 months, the pearl bud induction rate reaches 100%, and the average pearl bud formation number of a single mature branch is more than 20. The tamariskoid spikemoss herb after field planting is domesticated and cultured step by step in different cultivation environments such as a low-light greenhouse, a high-light greenhouse, an open field and the like, the field planting survival rate of the tamariskoid spikemoss herb can reach more than 93 percent, the blank of the artificial propagation technology of the tamariskoid spikemoss herb is filled, and the tamariskoid spikemoss herb has important significance for protecting wild species and developing and utilizing resources of the revived plant tamariskoid spikemoss herb.)

1. A bulbil propagation method for reviving selaginella pulvinata maxim is characterized by comprising the following steps:

(1) induction culture of Selaginella pulvinata's bulbil

Planting 2-3 years old plants of wild selaginella pulvinata maxim in a flowerpot filled with a matrix for pearl bud induction culture, wherein 2-3 kg/m of the matrix is added³The slow release fertilizer; the flowerpot planted with the cushion-shaped selaginella tamariscina plant is placed in a low-light-intensity greenhouse for cultivation, and the low-light-intensity greenhouse is controlled to be in: the temperature is 10-30 ℃, and the illumination intensity at noon on a sunny day is 150-180 mu mol.m^-2·s^-1The illumination intensity at noon in cloudy days and at noon in rainy days is 40-60 mu mol.m^-2·s^-1(ii) a Watering for 1 time every 4-5 days, and keeping the water content of the substrate to be 40-45%; pouring a compound fertilizer solution with the concentration of 0.100-0.125% w/v once every 2 weeks, wherein the compound fertilizer solution is water and N15% w/w, P₂O₅15％w/w，K₂Preparing O15% w/w compound fertilizer; the slow release fertilizer contains N14% and P in percentage by mass₂O₅14％，K₂O 14％，S 4％；

(2) Plug seedling culture of cushion-shaped selaginella tamariscina bulbels

After the cushion-shaped selaginella tamariscina bulbels obtained by induction in the step (1) begin to form more than 2 branches and more than 2 roots, separating the bulbels from the plants by using forceps, planting the bulbels in a plug tray filled with a substrate for plug seedling culture, and planting the plug tray with the cushion-shaped selaginella tamariscina bulbelsPlacing the mixture in the low-light greenhouse in the step (1) for culturing, wherein 1 time of water permeation is sprayed every 2 days from 1 week to 2 weeks of culture, and 1 time of water permeation is sprayed every 4-6 days from 3 weeks to 4 weeks; after 1 month, spraying a compound fertilizer solution with the concentration of 0.100-0.125% w/v for 1 time every 2 weeks, wherein the compound fertilizer solution is water and N15% w/w, P₂O₅15％w/w，K₂Preparing O15% w/w compound fertilizer;

(3) planting of cushion-shaped herba selaginellae bulbil plug seedling

When the crown width of the cushion-shaped herba selaginellae bulbil plug seedling in the step (2) reaches more than 2.0cm, fixedly planting the cushion-shaped herba selaginellae bulbil plug seedling in a flowerpot filled with a matrix, wherein 2-3 kg/m of the matrix is added³The slow release fertilizer comprises N14 percent and P by mass fraction₂O₅14％，K₂O 14％，S 4％；

(4) Cultivation management after field planting

In the 1 st month after field planting, the flowerpot is placed in the low-light greenhouse in the step (1) for field planting initial culture, and 1 time of water permeation is poured every 7-9 days; after culturing in a low-light greenhouse for 1 month, placing the flowerpot in a high-light greenhouse for culturing, and controlling the high-light greenhouse to: the temperature is 10-30 ℃, and the illumination intensity at noon on a sunny day is 1500-1700 mu mol.m^-2·s^-1The illumination intensity at noon in cloudy and rainy days is 400-500 [ mu ] mol.m^-2·s^-1Watering for 1 time per 5-6 days to permeate water, and watering a compound fertilizer solution with the concentration of 0.100-0.125% w/v once per 2 weeks, wherein the compound fertilizer solution is water and N15% w/w and P₂O₅15％w/w，K₂Mixing and preparing O15% w/w compound fertilizer; after culturing for 2-3 months in a strong illumination greenhouse, moving the flowerpot to an unshielded open-air field outside the greenhouse for open-air culture, watering for 1 time per 3-5 days, and watering a compound fertilizer solution with the concentration of 0.100-0.125% w/v once per 2 weeks, wherein the compound fertilizer solution is water and a compound fertilizer solution containing N15% w/w and P₂O₅15％w/w，K₂Mixed with compound fertilizer of O15% w/w.

2. The method for propagating the bulbil of the revived plant Selaginella pulvinata as claimed in claim 1, wherein:

the matrix in the step (1), the step (2) and the step (3) is a mixed matrix formed by mixing turf and perlite.

3. The method for propagating the bulbil of the revived plant Selaginella pulvinata as claimed in claim 2, wherein:

the volume ratio of the turf to the perlite in the mixed matrix in the step (1) is 8-10: 1; the volume ratio of the turf to the perlite in the mixed matrix in the step (2) and the step (3) is 4-5: 1.

4. The method for propagating the bulbil of the revived plant Selaginella pulvinata as claimed in claim 1, wherein:

in the step (1) and the step (4), the amount of the compound fertilizer solution with the concentration of 0.100-0.125% w/v is poured once every 2 weeks, and the mixed substrate in the flowerpot is fully poured; in the step (2), the amount of the compound fertilizer solution with the concentration of 0.100-0.125% w/v poured once every 2 weeks is that the mixed matrix in the hole tray is sprayed thoroughly.

5. The method for propagating the bulbil of the revived plant Selaginella pulvinata as claimed in claim 1, wherein:

the inner diameter of the flowerpot in the step (1) is 15-16 cm; in the step (2), the specification of the plug tray is 72 holes, and 1 bulbil is planted in each hole; and (4) in the step (3), the inner diameter of the flowerpot is 12-13 cm.

Technical Field

The invention belongs to the field of plant propagation, and particularly relates to a bulbil propagation method for reviving selaginella pulvinata maxim.

Background

Selaginella pulvinata (Hook. & Grev) Maxim.) belongs to Selaginella of Selaginellaceae, and is in cushion shape, and main stem is not in creeping shape, and mainly grows on the surface of exposed limestone or stone cracks, and the growing environment is dry. Selaginella pulvinata is commonly called as 'Jiu died hempleaf groundsel herb', is a typical resuscitation plant, has extremely strong dehydration resistance, can keep long-term survival through lower metabolism under the condition of extreme dehydration, and can quickly recover normal growth when the water condition is proper. Selaginella pulvinata is a special ornamental plant resource due to the miraculous resuscitation characteristic. Meanwhile, the selaginella pulvinata maxim contains medicinal components such as flavone, biflavone, alkynol and the like, has the effects of resisting cancer, viruses, oxidation, sugar reduction and the like, and is an important medicinal plant resource.

Selaginella pulvinata is an allosporidium fern, has megaspores and microspores, in a humid environment, the megaspores develop into female gametophytes with a cervicales, the microspores develop into male gametophytes with spermatids, in a water environment, the sperms in the spermatids enter the cervicales to be combined with ova to form fertilized ova, and finally, Selaginella pulvinata seedlings develop. However, Selaginella pulvinata is arid in the habitat, has a short humid period only in rainy season, is not beneficial to spore propagation of Selaginella pulvinata, and is difficult to form spore propagation offspring. Continuous field investigation finds that the natural reproduction rate of the wild population of the selaginella pulvinata maxim is low, and the number of individuals in the population is not obviously increased. In addition, the selaginella pulvinata maxim has ornamental and medicinal values, wild resources are greatly mined, so that the field population quantity of the selaginella pulvinata maxim is reduced sharply, and the wild resources cannot meet the current requirements.

In Selaginella, part of the main stem is of creeping type, such as: the small emerald green clouds have adventitious roots at the stem nodes and can be used for cuttage propagation, but the main stem of the selaginella pulvinata maxim is not creeping and is extremely difficult to perform cuttage propagation. In addition, in a wild habitat, the incidence rate of the bead buds of the selaginella pulvinata maxim is extremely low, and wild bead bud propagating plants are extremely few. At present, no report about the related propagation technology of the revived plant selaginella pulvinata or a pearl bud propagation method of the revived plant selaginella pulvinata is seen.

Disclosure of Invention

The invention provides a bulbil propagation method of resuscitated plant selaginella pulvinata, aiming at solving the technical problems of rapid reduction of wild population of the resuscitated plant selaginella pulvinata, low natural fertility and extremely difficult cuttage propagation.

The invention provides a method for reproducing the bead buds of resuscitation plant selaginella pulvinata which comprises the following steps:

(1) induction culture of Selaginella pulvinata's bulbil

Planting 2-3 years old plants of wild selaginella pulvinata maxim in a flowerpot filled with a matrix for pearl bud induction culture, wherein 2-3 kg/m of the matrix is added³The slow release fertilizer; the flowerpot planted with the cushion-shaped selaginella tamariscina plant is placed in a low-light-intensity greenhouse for cultivation, and the low-light-intensity greenhouse is controlled to be in: the temperature is 10-30 ℃, and the illumination intensity at noon on a sunny day is 150-180 mu mol.m^-2·s^-1The illumination intensity at noon in cloudy days and at noon in rainy days is 40-60 mu mol.m^-2·s^-1(ii) a Watering for 1 time every 4-5 days, and keeping the water content of the substrate to be 40-45%; pouring a compound fertilizer solution with the concentration of 0.100-0.125% w/v once every 2 weeks, wherein the compound fertilizer solution is water and a compound fertilizer solution containing N15% w/w, P₂O₅15％w/w，K₂Mixing and preparing O15% w/w compound fertilizer according to the concentration; the slow release fertilizer contains N14% and P in percentage by mass₂O₅14％，K₂O 14％，S 4％；

(2) Plug seedling culture of cushion-shaped selaginella tamariscina bulbels

After the cushion-shaped selaginella tamariscina bulbels obtained by induction in the step (1) begin to form more than 2 branches and more than 2 roots, separating the selaginella tamariscina bulbels from the plants by using forceps and planting the selaginella tamariscina bulbels in a plug tray filled with a substrate for plug seedling culture, and planting the plug tray filled with the cushion-shaped selaginella tamariscina bulbelsPlacing the mixture in the low-light greenhouse in the step (1) for culturing, wherein 1 time of water permeation is sprayed every 2 days from 1 week to 2 weeks of culture, and 1 time of water permeation is sprayed every 4-6 days from 3 weeks to 4 weeks; after 1 month, spraying a compound fertilizer solution with the concentration of 0.100-0.125% w/v for 1 time every 2 weeks, wherein the compound fertilizer solution is water and N15% w/w, P₂O₅15％w/w，K₂Mixing and preparing O15% w/w compound fertilizer according to the concentration;

(3) planting of cushion-shaped herba selaginellae bulbil plug seedling

When the crown width of the cushion-shaped herba selaginellae bulbil plug seedling in the step (2) reaches more than 2.0cm, fixedly planting the cushion-shaped herba selaginellae bulbil plug seedling in a flowerpot filled with a matrix, wherein 2-3 kg/m of the matrix is added³The slow release fertilizer contains N14% and P in percentage by mass₂O₅14％，K₂O 14％，S 4％；

(4) Cultivation management after field planting

In the 1 st month after field planting, the flowerpot is placed in the low-light greenhouse in the step (1) for field planting initial culture, and 1 time of water permeation is poured every 7-9 days; after culturing in a low-light greenhouse for 1 month, placing the flowerpot in a high-light greenhouse for culturing, and controlling the high-light greenhouse to: the temperature is 10-30 ℃, and the illumination intensity at noon on a sunny day is 1500-1700 mu mol.m^-2·s^-1The illumination intensity at noon in cloudy and rainy days is 400-500 [ mu ] mol.m^-2·s^-1Watering for 1 time per 5-6 days to permeate water, and watering a compound fertilizer solution with the concentration of 0.100-0.125% w/v once per 2 weeks, wherein the compound fertilizer solution is water and N15% w/w and P₂O₅15％w/w，K₂Mixing and preparing O15% w/w compound fertilizer according to the concentration; after culturing for 2-3 months in a strong illumination greenhouse, moving the flowerpot to an open-air place without shielding outside the greenhouse for open-air culture, watering for 1 time per 3-5 days, and watering a compound fertilizer solution with the concentration of 0.100-0.125% w/v once per 2 weeks, wherein the compound fertilizer solution is water and a compound fertilizer solution containing N15% w/w and P₂O₅15％w/w，K₂And mixing and preparing the O15% w/w compound fertilizer according to the concentration.

Further, the matrix in the step (1), the step (2) and the step (3) is a mixed matrix formed by mixing turf and perlite.

Further, the volume ratio of the turf to the perlite in the mixed matrix in the step (1) is 8-10: 1; the volume ratio of the turf to the perlite in the mixed matrix in the step (2) and the step (3) is 4-5: 1.

Further, in the step (1) and the step (4), the amount of the compound fertilizer solution with the concentration of 0.100-0.125% w/v is poured once every 2 weeks, and the mixed substrate in the flowerpot is fully poured; in the step (2), the amount of the compound fertilizer solution with the concentration of 0.100-0.125% w/v poured once every 2 weeks is that the mixed matrix in the hole tray is sprayed thoroughly.

Further, the inner diameter of the flowerpot in the step (1) is 15-16 cm; in the step (2), the specification of the plug tray is 72 holes, and 1 bulbil is planted in each hole; and (4) in the step (3), the inner diameter of the flowerpot is 12-13 cm.

The invention has the main innovation points and beneficial effects that:

1. the invention successfully induces the formation of the bead buds of the selaginella pulvinata maxim.

The method screens out a culture medium, a lighting condition and a moisture condition which are suitable for the induction of the cushion-shaped selaginella tamariscina pearl buds, the induction rate of the pearl buds reaches 100 percent after 2-3-year-old plants of the wild cushion-shaped selaginella tamariscina are subjected to the pearl bud induction culture for 5-6 months, and the average pearl bud formation number of a single mature branch is more than 20 pearl buds.

2. The method obtains higher planting survival rate of the spikemoss herb bulbils and realizes the artificial propagation of the spikemoss herb.

The method carries out step-by-step domestication culture on the planted selaginella pulvinata bulbils in different culture environments such as a low-light greenhouse, a high-light greenhouse, an open field and the like, the planting survival rate of the selaginella pulvinata bulbils can reach more than 93 percent, and the blank of the selaginella pulvinata artificial propagation technology is filled.

Drawings

FIG. 1: and (4) inducing the generation of bulbels on the branches of the cushion-shaped selaginella tamariscina. The induced formation of the bulbils is indicated by the arrows in FIG. 1. The line segments in fig. 1 are to scale and have a length of 500 μm.

FIG. 2: the bulbil on a single shoot of selaginella pulvinata maxim begins to form a plantlet. In FIG. 2, the line segment is a scale and the length is 2 mm.

Detailed Description

The following examples facilitate a better understanding of the invention. The test methods in the following examples are conventional methods unless otherwise specified. The test materials used in the following examples are commercially available, and the results of the quantitative tests in the examples were averaged by setting three replicates.