Toxin type plant rooting solution, rooting method and application

Abstract

The present application relates to a kind of toxin type plant rooting liquid, rooting method and application, wherein the composition of rooting liquid is poplar canker bacteria toxin, formula is 5.00% poplar canker bacteria toxin aqueous solution, using method is the base of the stem section to be rooted into the rooting liquid, when the stem section base appears root primordium, transplant to soil, rooting rate can reach 83.33%, and more fibrous root, plant growth is robust, seedling survival rate can reach 83.33%, simultaneously, the rooting liquid has the ability of enhancing seedling disease resistance simultaneously.This application not only can efficiently promote plant rooting, improve transplanting survival rate, but also can improve the immunity of plant, low cost, simple to make, convenient to use, remarkable effect.

Description

Technical Field

[0001] This invention belongs to the field of plant rooting agent technology, and relates to a toxic plant rooting solution, a rooting method and its application. Background Technology

[0002] Forest ecosystems play a vital role in sustaining life on Earth. However, against the backdrop of current global climate change, various pest and disease disasters are occurring frequently and causing increasingly severe damage. Therefore, improving afforestation survival rates, enhancing tree resistance to biological stresses, and ultimately improving forest quality are important issues that need further resolution. Among these, cultivating resistant afforestation tree species or seedlings and developing effective resistant seedling cultivation techniques are key to solving this problem.

[0003] Branch cuttings are a common method of asexual plant propagation, widely used in forestry for afforestation of fast-growing plants such as poplar and willow, and also for the asexual propagation of economic crops and flower seedlings. Due to their ability to promote adventitious root formation and rooting of cuttings, plant growth regulators such as indoleacetic acid (IAA), indolebutyric acid (IBA), naphthaleneacetic acid (NAA), ethylene, gibberellin, and brassinosteroids are widely used in the cutting production of various forest trees. ABT rooting powder is the most widely used broad-spectrum and highly effective rooting promoter. While participating in the process of adventitious root formation, ABT rooting powder also supplements exogenous hormones and promotes the synthesis of endogenous hormones in plants, thus promoting adventitious root formation and shortening rooting time.

[0004] However, plant growth regulators such as ABT rooting powder do not specifically improve the resistance of plant seedlings to pests and diseases. While various mycorrhizal fungi and EM microbial preparations promote plant growth, there are no reports showing that these products can induce adventitious root formation. Furthermore, these preparations are usually made from symbiotic or probiotic bacteria and do not contain pathogenic factors or toxicity, thus they generally do not induce disease resistance in plants. Therefore, seedling cultivation techniques that simultaneously promote rooting and enhance resistance remain a crucial technology for afforestation that urgently needs to be addressed. Currently, there are no reports of using fungal toxins or bacterial toxins to promote plant rooting, nor are there any rooting products that simultaneously promote rooting and enhance disease resistance. Summary of the Invention

[0005] In view of this, the present invention provides a toxic plant rooting solution that promotes plant rooting and enhances disease resistance, its method of use and its application. It can not only promote plant rooting, accelerate the growth rate of new roots and increase the number of lateral roots, but also promote plant growth and enhance plant disease resistance.

[0006] To address the aforementioned technical problems, the present invention provides a rooting solution consisting of a 5.00% aqueous solution of poplar canker toxin. The method of application involves inserting the base of the stem segment to be rooted into the rooting solution. Once root primordia appear at the base of the stem segment, it is transplanted into the soil. The rooting rate can reach 83.33%, with robust root growth and numerous fibrous roots, resulting in a seedling survival rate of 83.33%. Simultaneously, this rooting solution activates a series of defense-related genes in the seedlings, enhancing their disease resistance.

[0007] This invention discloses a toxic plant rooting solution that promotes plant rooting and enhances plant disease resistance, comprising a rooting solution, wherein the rooting solution is an aqueous solution of poplar canker toxin at a concentration of 5.00%.

[0008] Furthermore, the poplar canker pathogen toxin is an acetone-precipitated pathogen toxin.

[0009] Furthermore, the method for extracting the aqueous solution of poplar canker pathogen toxin includes the following steps:

[0010] (1) The ulcer strain was activated on 2.0% PDA medium (2.0% potato extract, 2.0% glucose, 1.5% agar) and then cultured in the dark at 25°C.

[0011] (2) After the strain was cultured in PDA for 7 days, 5 bacterial cakes were inoculated in each bottle under aseptic conditions and cultured in the dark at 25℃, 110r / min for 10 days with shaking.

[0012] (3) After filtration of the cultured bacterial solution under sterile conditions, the filtrate was vacuum evaporated using a rotary evaporator at 38℃-40℃. Acetone was added, and the solution was shaken and extracted at room temperature for 1 hour to obtain the original bacterial toxin solution, which was then stored at 4℃ for later use.

[0013] (4) The toxin treatment solution is prepared at a volume ratio of 5.00%.

[0014] A method for promoting plant rooting and improving plant disease resistance involves inserting the base of a stem segment to be rooted into a rooting solution. Once root primordia appear at the base of the stem segment, the segment is transplanted into soil. The rooting solution contains a 5.00% aqueous solution of poplar canker toxin.

[0015] Furthermore, when the treated plant material is transplanted into a substrate such as soil, there is no need to disinfect the substrate beforehand.

[0016] The application of the toxin-type rooting solution that promotes both root growth and disease resistance in plants.

[0017] Compared with the prior art, the technical effects of the present invention include:

[0018] This invention can not only efficiently promote plant rooting and improve transplant survival rate, but also enhance plant immunity. It is low in cost, simple to make, easy to use, and has significant effects. Attached Figure Description

[0019] like Figure 1 As shown: Rooting status of Xinjiang poplar branches treated with 5.00% toxin solution (a) and control group treated with sterile water (b);

[0020] Figure 2 Growth of Xinjiang poplar branches treated with 5.00% toxin solution (a) and sterile water treatment control (b) after transplanting; Detailed Implementation

[0021] The present invention will be further described in detail below through embodiments, which are intended to illustrate the invention and not limit it. The scope of protection of the present invention should be determined by the scope defined in the claims. It should be noted that the preferred embodiments should not be regarded as a limitation of the present invention. For those skilled in the art, various improvements and modifications can be made to the present invention without departing from the principles of the invention, and these improvements and modifications also fall within the scope of protection of the present invention.

[0022] The invention will be further illustrated below with examples. The plant example mainly uses *Populus alba var. pyramidalis* Bge., and the fungus example mainly uses *Botryosphaeria dothidea* strain CZA. This does not limit the scope of the invention.

[0023] 1. CZA plate culture of ulcer pathogens

[0024] The fungal material was the ulcer disease strain CZA preserved by the Water Physiology Research Laboratory of the Chinese Academy of Forestry. After activation on 2.0% PDA medium (2.0% potato extract, 2.0% glucose, 1.5% agar), strain CZA was cultured in the dark at 25°C.

[0025] 2. Liquid culture of CZA bacteria causing ulceration

[0026] After 7 days of PDA culture, the strain was aseptically inoculated into 500 ml Erlenmeyer flasks containing 300 ml of sterilized potato glucose broth (PD) with 5 pieces of mycelium per flask. The flasks were then inoculated with shaking for 10 days at 25°C, 110 r / min and in the dark.

[0027] 3. Extraction of CZA toxin from Ulcerative Colitis Bacteria

[0028] The cultured bacterial culture was filtered under aseptic conditions using a 0.2 μm microporous membrane. The resulting filtrate was then vacuum evaporated to 30 mL using a rotary evaporator at 38-40 °C. 150 mL of acetone was added in three portions, and the mixture was extracted with shaking at room temperature for 1 hour. The acetone was then removed, and the remaining insoluble fraction was vacuum evaporated to remove the acetone. The fraction was then dissolved in sterile water and brought to a final volume of 300 mL to obtain the stock solution of the bacterial toxin, which was stored at 4 °C for later use.

[0029] 4. Treatment of Xinjiang poplar and Beijing poplar stem segments with CZA toxin from the ulcer pathogen.

[0030] Prepare toxin treatment solutions (aqueous solutions) according to volume ratios of 10%, 5%, 2.50%, and 1.25%, respectively. Use sterile water as a control. Use approximately 400ml plastic culture bottles as treatment containers, with each bottle containing 3cm of toxin treatment solution. Cut one-year-old poplar branches into 15cm stem segments, inserting groups of three stem segments into the toxin treatment solution. Place the bottles in a 25℃ light incubator, changing the toxin dilution solution daily.

[0031] 5. Greenhouse transplanting

[0032] When root primordia appear at the lower end of poplar stem segments, the segments are removed from the toxin-treated solution. Some segments are transferred to culture bottles filled with tap water (10-12 cm high) and cultured in a 25℃ light incubator, with the tap water changed daily. After 30 days, the number of rooted stem segments is counted, and the length of the tap root is measured. Some stem segments are directly planted in a culture medium (peat moss: perlite = 6:1), watered with tap water, and placed directly in a greenhouse for cultivation without covering with plastic film. Routine seedling management is then carried out, and the survival rate and seedling growth are observed after 10 days (Table 1).

[0033] Table 1. Rooting and survival rates of poplar stem segments transplanted in greenhouse under different concentrations of poplar canker fungal toxin treatment.

[0034]

[0035]

[0036] Table 1. Rooting and survival rates of poplar stem segments from Xinjiang after treatment with different concentrations of poplar bark rot virulence solution and greenhouse transplanting.

[0037]

[0038] 6. Detection of disease resistance gene expression

[0039] Poplar stem segments treated with 0.00% and 5.00% toxin were inoculated with *Populus canker* using the hole-punching inoculation method. Fully expanded leaves and bark 2 cm from the inoculation hole were collected, and total RNA was extracted from the leaves and bark, respectively. The RNA was reverse transcribed into cDNA, and the expression of two NBS-LRR genes (Potri.006G271600 and Potri.008G212200) was quantitatively analyzed using gene-specific primers (Table 1). The results showed that compared with the control treated with sterile water, the expression of NBS-LRR resistance genes in poplar leaves and bark treated with 5.00% toxin was significantly increased (e.g., ...). Figure 1 As shown in the figure, this indicates that treatment with the toxin solution can increase the expression of disease-resistant genes in poplar trees, thus enhancing the disease resistance of seedlings.

[0040] Table 1 Primers for real-time quantitative PCR of disease resistance genes and internal reference genes.

[0041]

[0042]

[0043] The above description is only a preferred embodiment of the present invention, but the scope of protection of the present invention is not limited thereto. Any equivalent substitutions or modifications made by those skilled in the art within the scope of the technology disclosed in the present invention, based on the technical solution and inventive concept of the present invention, should be covered within the scope of protection of the present invention.

Claims

1. A toxic plant rooting solution, comprising a rooting solution, characterized in that, The rooting solution is a 5.00% aqueous solution of poplar canker pathogen toxin; the poplar canker pathogen toxin is an acetone-precipitated pathogen toxin; the poplar canker pathogen toxin is prepared by culturing Botryosphaeria dothidea strain CZA.

2. The toxic plant rooting solution according to claim 1, characterized in that, The method for extracting the toxin aqueous solution from the poplar canker pathogen includes the following steps: (1) The Botryosphaeria dothidea strain CZA described in claim 1 was activated on 2.0% PDA medium and then cultured in the dark at 25°C; (2) After the strain was cultured in PDA for 7 days, 5 bacterial cakes were inoculated into each bottle under aseptic conditions and cultured in the dark at 25℃, 110r / min for 10 days with shaking. (3) After filtration under sterile conditions, the filtrate was vacuum evaporated at 38℃-40℃ using a rotary evaporator. Acetone was added, and the mixture was shaken and extracted at room temperature for 1 hour to obtain the original bacterial toxin solution, which was then stored at 4℃ for later use. (4) Prepare a 5.00% aqueous solution of poplar canker pathogen toxin by mixing the original stock solution of the pathogen toxin with the volume ratio.

3. A method for promoting plant rooting and enhancing plant disease resistance using the toxic plant rooting solution described in claim 1, characterized in that, Insert the base of the stem segment to be rooted into the rooting solution. When root primordia appear at the base of the stem segment, transplant it into the soil. The rooting solution contains an aqueous solution of poplar canker toxin as described in claim 1 at a concentration of 5.00%.

4. The method according to claim 3, characterized in that: When the treated plant material is transplanted into the soil substrate, there is no need to disinfect the soil substrate beforehand.

5. The application of the toxic rooting solution according to claim 1 in promoting plant rooting and improving plant disease resistance.

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