Tung oil meal, tung oil meal water extract and use thereof in reducing banana wilt disease incidence and promoting banana growth

Abstract

The present invention provides a tung oil meal, a tung oil meal water extract and an application thereof in reducing the incidence of banana wilt disease and promoting banana growth. The tung oil meal and the tung oil meal water extract of the present invention can effectively inhibit the growth of banana wilt disease No. 4 physiological subspecies (Foc 4) in the banana cultivation process, reduce the incidence of banana wilt disease in the seedling stage, and at the same time, during the growth of banana seedlings, stimulate the activity of root polyphenol oxidase (POD), superoxide dismutase (SOD), catalase (CAT), polyphenol oxidase (PPO) and other cell protection enzymes to increase their content, thereby stimulating root growth, especially root meristem growth, and can promote leaf growth, increase chlorophyll content, increase dry matter accumulation, etc. through the growth and development of the root system, thereby achieving effective promotion of growth and development, and has broad application prospects in the industry.

Description

Technical Field

[0001] The invention relates to the technical field of banana cultivation, and in particular to tung oil meal, tung oil meal water extract and application thereof in reducing the incidence of banana wilt disease and promoting banana growth. Background Art

[0002] Banana wilt is a typical soil-borne disease. Its Fusarium oxysporum f.sp. cubense 4 (Foc 4) has caused a devastating blow to the world's banana industry. The prevention and control of banana wilt has been a top priority in the industry and research in recent decades. Existing research reports and production practices have shown that the selection and breeding of new banana wilt-resistant varieties and green integrated prevention and control measures are more effective and are the current key development direction. Among the cultivation measures, water-dry crop rotation, intercropping of leeks, increased application of organic fertilizers, regular application of biological fertilizers, and strict management of entry and exit can effectively prevent and reduce the incidence of banana wilt. At a time when labor costs and production costs are rising and the average per-acre benefit of banana orchards is declining, there is still a strong demand for convenient, low-cost, and efficient banana wilt prevention and control measures.

[0003] The utilization and development of natural wastes such as cakes and meal is not only related to reducing environmental pollution, but also an important way to improve economic benefits. They are usually used for secondary utilization as feed additives, fertilizers, etc., and are also raw materials for the extraction of organic compounds. Rapeseed meal, cottonseed meal, etc. are also widely used in banana cultivation, and have significant functions in improving fertility. Tung tree is a perennial woody tree, and tung oil is an industrial vegetable oil made by mechanically pressing and refining the picked tung fruit. As an important oil product in my country, tung oil produces a large amount of tung oil meal every year, but the application of tung oil meal in banana cultivation has not been reported. Summary of the invention

[0004] In view of this, the present invention provides a tung oil meal, a tung oil meal water extract and applications thereof in reducing the incidence of banana wilt disease and promoting banana growth.

[0005] The technical solution of the present invention is achieved in this way:

[0006] Application of tung oil meal in banana cultivation.

[0007] Furthermore, the tung oil cake is used to reduce the incidence of banana wilt disease and promote banana growth.

[0008] Furthermore, the tung oil cake is used in preparing banana cultivation fertilizer or regulator.

[0009] Furthermore, the tung oil cake is used in the preparation of a fertilizer or regulator for inhibiting Foc 4 banana wilt disease.

[0010] Furthermore, the tung oil cake is used in the preparation of a fertilizer or regulator for increasing the POD, SOD, CAT, and PPO activity of banana roots.

[0011] The tung oil meal water extract is prepared from the tung oil meal.

[0012] Furthermore, the preparation method of the tung oil meal water extract is: weigh 8-10g of tung oil meal, add it into 100-120mL of sterile water, put it into a shaker at 100-120rpm, extract it at 26-30℃ for 22-26h, and filter it to obtain the tung oil meal water extract.

[0013] Furthermore, the preparation method of the tung oil meal water extract is: weigh 10 g of tung oil meal, add it into 100 mL of sterile water, put it into a shaker at 100 rpm, extract it at 28° C. for 24 hours, and filter it to obtain the tung oil meal water extract.

[0014] Furthermore, the tung oil cake water extract is used to reduce the incidence of banana wilt and promote banana growth.

[0015] Furthermore, the tung oil cake water extract is used in the preparation of a fertilizer or regulator for inhibiting Foc 4 banana wilt disease.

[0016] Furthermore, the tung oil cake water extract is used in the preparation of a fertilizer or regulator for increasing the POD, SOD, CAT, and PPO activity of banana roots.

[0017] Furthermore, the fertilizer is organic fertilizer.

[0018] Compared with the prior art, the present invention has the following beneficial effects:

[0019] The present invention applies tung oil cake and tung oil cake water extract prepared from the tung oil cake to banana cultivation. The tung oil cake and tung oil cake water extract of the present invention can effectively inhibit the growth of banana wilt disease No. 4 physiological subspecies (Foc 4) during banana cultivation, reduce the incidence of banana wilt disease in the seedling stage, and at the same time, during the growth of banana seedlings, stimulate the activity of root polyphenol oxidase (POD), superoxide dismutase (SOD), catalase (CAT), polyphenol oxidase (PPO) and other cell protection enzymes to increase their content, thereby stimulating root growth, especially root meristem growth, and can promote leaf growth, increase chlorophyll content, increase dry matter accumulation, etc. through the growth and development of the root system, thereby achieving effective promotion of growth and development, and has broad application prospects in the industry.

[0020] The present invention can extract active substances from tung oil cake by adopting a suitable water extraction process, thereby improving the ability of the tung oil cake to inhibit the growth of banana Fusarium wilt race 4 (Foc 4), reducing the incidence of Fusarium wilt at the seedling stage, and also promoting the growth and development of banana root system. BRIEF DESCRIPTION OF THE DRAWINGS

[0021] Figure 1 The effect of adding different proportions of tung oil cake to the cultivation medium on the resistance of banana seedlings to banana wilt disease; in the figure, Figure 1 a is the phenotypic observation of the incidence of banana seedlings, where A is the banana seedlings of the negative control group of group 1, C is the banana seedlings of the positive control group of group 2, E is the banana seedlings of group 3, G is the banana seedlings of group 4, and I is the banana seedlings of group 5; Figure 1 b is the observation of the disease index of banana seedlings, wherein B is the anatomical diagram of the rhizome of banana seedlings in the negative control group of group 1, D is the anatomical diagram of the rhizome of banana seedlings in the positive control group of group 2, F is the anatomical diagram of the rhizome of banana seedlings in group 3, H is the anatomical diagram of the rhizome of banana seedlings in group 4, and J is the anatomical diagram of the rhizome of banana seedlings in group 5.

[0022] Figure 2 Statistics on the effects of adding different proportions of tung oil cake to the cultivation medium on the internal and external morbidity indexes of Fusarium wilt in banana seedlings.

[0023] Figure 3 The figures show the effect of different dilution multiples on the inhibition of Foc 4 by the water extract of tung oil meal; in the figure, A is the growth of plate culture colonies in group A, B is the growth of plate culture colonies in group B, C is the growth of plate culture colonies in group C, D is the growth of plate culture colonies in group D, E is the growth of plate culture colonies in group E, and F is the inhibition rate of each group; in the figure, different letters abc indicate significant differences (P<0.05). DETAILED DESCRIPTION

[0024] In order to better understand the technical content of the present invention, specific embodiments are provided below to further illustrate the present invention.

[0025] Unless otherwise specified, the experimental methods used in the embodiments of the present invention are all conventional methods.

[0026] Unless otherwise specified, the materials, reagents, etc. used in the embodiments of the present invention can be obtained from commercial sources.

[0027] The banana variety used for the test was Musa Cavendish subgroup cv brazil (AAA). The five-leaf, one-heart cup seedlings of Musa Cavendish were purchased from Hainan Tropical Crops and Two Institutes Seed Technology Co., Ltd.; tung oil meal was purchased from Gucheng County, Xiangyang City, Hubei Province; and Fusarium oxysporum f.sp. cubense 4 (Foc 4) was provided by the Institute of Environment and Plant Protection, Chinese Academy of Tropical Agricultural Sciences.

[0028] Example 1 - Effect of tung oil meal on the incidence of banana wilt

[0029] In the same environment, a pot experiment was conducted with Brazilian banana seedlings at the five-leaf and one-heart stage as the research material. During the experiment, the experiment was divided into five groups 1-5, with 9 pots in each group. Groups 1-5 added tung oil cake as a regulator at 0g / L, 0g / L, 10g / L, 1g / L, and 0.1g / L to the cultivation matrix and mixed evenly (the mass ratio of nutrient soil to vermiculite in the cultivation matrix was = 2:1). The mixed cultivation matrix was put into small pots, with 1L of soil per pot, and then banana seedlings were planted. One week later, the Foc 4 bacterial suspension with a concentration of 105 spores / ml was irrigated to groups 2-5 at a volume of 100mL / by the root injury inoculation method. Group 1 was irrigated with 100mL of clean water as the clean water control group, and Group 2 was the positive control group. After regular observation, the incidence rate and internal and external disease index of banana seedlings in each group were counted 4 weeks after the banana seedlings were planted. External symptoms: banana seedlings have yellow spots on leaves, petioles bend and droop, leaves wither or the entire seedling dies. Internal symptoms: yellow or reddish-brown spots can be seen between the pith and cortex of the central column when the rhizome is cut, or the root turns dark brown and dries up after cutting. The results are as follows Figure 1 shown.

[0030] as follows Figure 1 As shown, from the perspective of morbidity, all the positive groups in group 2 were diseased, and the morbidity of banana seedlings in groups 3-5 were 66.67%, 73.33% and 80%, respectively, which were all lower than the 100% morbidity of the positive control. From the perspective of disease index, banana seedlings in groups 3-5 had lower internal disease index and external disease index. The internal disease index and external disease index of banana seedlings in group 3 were 5.53 and 3.33, respectively, the internal disease index and external disease index of banana seedlings in group 4 were 6.66 and 3.8, respectively, and the internal disease index and external disease index of banana seedlings in group 5 were 6.93 and 4, respectively, which were lower than the internal disease index of 7.73 and the external disease index of 4.2 of the positive control in group 2. This shows that the addition of tung oil meal to the cultivation medium can effectively reduce the incidence of bananas.

[0031] Example 2 - Effect of tung oil cake on enzyme activity in roots of banana seedlings

[0032] In the same environment, a potted experiment was carried out with five-leaf and one-heart Brazilian banana seedlings as the research material. During the experiment, it was divided into five groups, 1-4, with 9 pots in each group. For groups 1-4, tung oil cake was added as a regulator at 0g / L, 10g / L, 1g / L, and 0.1g / L to the cultivation medium and mixed evenly (the mass ratio of nutrient soil: vermiculite in the cultivation medium was 2:1). The mixed cultivation medium was put into small pots with 1L of soil in each pot, and then banana seedlings were planted. Group 1 was sprayed with 0.3% urea aqueous solution once a week as the control group, and spraying was used as the standard. Groups 2-4 were sprayed with the same amount of clean water every week. One week after the banana seedlings were planted, the Foc 4 suspension with a concentration of 105 spores / ml was watered at a volume of 100 mL / min by the root wound inoculation method. Five days later, the activities of peroxidase (POD), catalase (CAT), phenylalanine ammonia lyase (PAL), polyphenol oxidase (PPO), and superoxide dismutase (SOD) in the roots of the banana seedlings of each group were measured. The determination method was based on the kit (Solarbio, peroxidase (POD) activity detection kit; Solarbio, catalase (CAT) activity detection kit; Solarbio, phenylalanine ammonia lyase (PAL) activity detection kit). The results are shown in Table 1.

[0033] Table 1: Effects of adding tung oil cake to the cultivation medium on the activities of five antioxidant enzymes in the roots of banana seedlings

[0034]

[0035] Note: Different superscript letters in the same column indicate significant differences (P<0.05).

[0036] Peroxidases in many plants are often used as physiological indicators and marker genes for disease resistance. Antioxidant enzymes such as POD, SOD, CAT, PPO and PAL together constitute cell protection enzymes, which play an important role in maintaining normal cell metabolism. The above results show that after adding tung oil cake to the seedling medium, the detection of peroxidase activity in the roots of banana seedlings found that after inoculation with Foc 4, the activity of POD, SOD, CAT and PPO in the roots of seedlings could be stimulated; and after adding tung oil cake, the activity of POD, SOD, CAT and PAL would be further improved, among which the difference in the activity of POD, SOD and PPO was significant.

[0037] Example 3 - Effect of tung oil meal on the growth and development of banana seedlings

[0038] Under the same environment, a potted experiment was conducted with five-leaf and one-heart Brazilian banana seedlings as research materials. During the experiment, the experiment was divided into five groups 1-4, with 9 pots in each group. Groups 1-4 added tung oil cake as fertilizer to the cultivation matrix at 0g / L, 10g / L, 1g / L, and 0.1g / L respectively (the mass ratio of nutrient soil: vermiculite in the cultivation matrix was = 2:1), and the mixed cultivation matrix was put into small pots, with 1L of soil in each pot, and then banana seedlings were planted. Group 1 was sprayed with a 0.3% urea aqueous solution once a week as a control group, and the spraying was based on wetness. Groups 2-4 were sprayed with the same amount of water every week. After the banana seedlings grew for 4 weeks, the changes in fresh weight, dry weight, chlorophyll content, root length, root volume, root surface area, and root diameter of the seedlings in each group were detected. The results are shown in Table 2 below.

[0039] Table 2 Statistics on the promotion of seedling growth and development by adding different proportions of tung oil meal

[0040]

[0041] Note: Different superscript letters in the same column indicate significant differences (P<0.05).

[0042] The above results show that compared with the control group 1 which used chemical fertilizers conventionally, after using tung oil meal, the banana leaves were dark green and thick, and the dry weight and fresh weight of the leaves were significantly higher than the control group. The chlorophyll content was higher than the control group, but the difference between the 4th group with tung oil meal added at 0.1g / L and the control group was not obvious; further observation of the root system showed that the root system of the treatment group with tung oil meal added was more developed. By measuring the root system of Brazilian banana seedlings in groups 2-4, it was found that the root length, root volume, and root surface area were all higher than those in the control group, among which the treatment group 3 was the highest and significantly higher than other treatments. Specifically, the root length, root volume, and root surface area were the highest in the treatment group 3, which were significantly higher than those in the control group, but the root diameter showed that the treatment group was close to the control group except for the 2nd group, and the other two groups were significantly lower than the control group. It shows that the addition of tung oil meal to the cultivation medium can promote the growth of the root system of banana seedlings, and the development of the root system can promote leaf growth, increase chlorophyll content, and accumulate dry matter, thereby achieving the effect of promoting the growth and development of banana seedlings.

[0043] Example 4 - Effect of Tung Oil Meal Water Extract on Foc 4 Inhibition

[0044] The inhibitory effect of tung oil cake extract on Foc 4 was studied by plate inhibition experiment, and the growth process of mycelium inhibition and the influence of different dilution multiples on the inhibitory effect were observed.

[0045] Preparation method of tung oil meal water extract: weigh 10g of tung oil meal, add it into 100mL of sterile water, put it into a shaker at 100rpm, extract it at 28℃ for 24h, and filter it to obtain the tung oil meal water extract.

[0046] The tung oil cake water extract prepared in the above manner was added to the sterilized PDA medium at 10 μL / mL in four gradients of undiluted, 10-fold, 100-fold, and 1000-fold dilution, corresponding to Group B, Group C, Group D, and Group E, respectively. After shaking well, it was poured into an 85 mm bacterial culture dish at 20 mL / dish. The plate without the addition of tung oil cake water extract was used as the control for Group A. After the plate solidified, a circular Foc 4 bacterial block with a diameter of 0.5 cm was inoculated and placed in an incubator at 28°C for 7 days. The colony diameter was measured and recorded every day, and the antibacterial rate (%) was calculated. The results are as follows Figure 3 shown.

[0047] Inhibition rate = (control colony diameter - treated colony diameter) / control colony diameter × 100%)

[0048] like Figure 3 As shown in the figure, the results showed that the tung oil cake water extract had a significant inhibitory effect on the growth of banana wilt pathogen. The colony diameter of the undiluted tung oil cake water extract in group B was significantly different from that of the control after being added to the plate. The inhibition rate increased from the second day to the fourth day, reaching the highest inhibition rate, and then gradually decreased. The inhibition rates from 2 to 7 days were 52.6%, 59.52%, 56.54%, 53.73%, 51.9%, and 49.42%, respectively. Tung oil cake can effectively inhibit the growth of Foc 4 mycelium. The inhibition rate increased from the second day to the fourth day, reaching the highest inhibition rate, and then gradually decreased. After the tung oil cake water extract was diluted at different times, the inhibition rate of the tung oil cake water extract was significantly reduced. There was no significant difference between the dilution of 10 times and the dilution of 100 times, but there was a significant difference between the dilution of 100 times and the dilution of 1000 times. After the C group was diluted 10 times, the inhibition rates from 2 to 7 days were 36.15%, 39.17%, 38.66%, 38.38%, 38.28%, and 37.05%, respectively. After the D group was diluted 100 times, the inhibition rates from 2 to 7 days were 34.79%, 37.20%, 38.66%, 38.09%, 37.42%, and 36.72%, respectively. After the E group was diluted 1000 times, the inhibition rates from 2 to 7 days were 19.19%, 26.21%, 27.58%, 29.16%, 30.28%, and 30.29%, respectively.

[0049] The tung oil cake water extract was diluted 10 times, 100 times, and 1000 times as fertilizer to irrigate banana seedlings once a week, 100 mL each time, and then the same amount of 0.3% urea aqueous solution was sprayed as the control group. After 4 weeks of planting, the root system of the banana seedlings was measured and it was found that the root length, root volume, and root surface area of ​​the banana seedlings irrigated with tung oil cake water extract were also higher than those of the control group. In addition, the fresh weight, dry weight, and chlorophyll content of the banana seedlings irrigated with tung oil cake water extract were also higher than those of the control group. This shows that tung oil cake water extract can promote the growth of banana seedlings.

[0050] In addition, tung oil meal ethanol extract was prepared. The preparation method was as follows: 10 g of tung oil meal was weighed and added to 100 mL of anhydrous ethanol, placed in a shaker at 100 rpm, extracted at 28 ° C for 24 hours, and filtered to obtain tung oil meal ethanol extract. The antibacterial effect of the undiluted tung oil meal ethanol extract on Foc 4 bacteria was determined. The antibacterial experimental method was the same as that of the tung oil meal ethanol extract. The antibacterial rates from 2 to 7 days were 39.2%, 51.38%, 46.37%, 45.63%, 46.49%, and 35.61%, respectively. Although the tung oil meal ethanol extract can also inhibit the growth of Foc 4 mycelium, its antibacterial effect is significantly lower than that of the tung oil meal water extract. Furthermore, the tung oil meal ethanol extract was diluted 10 times, 100 times, and 1000 times with water and then watered banana seedlings in the same way. The indicators of promoting banana growth were worse than those of the tung oil meal water extract with the same dilution multiple.

[0051] In addition, through the study of tung oil cake water extract, it was found that substances that can play an antibacterial role contain water-soluble substances such as protein macromolecules.

[0052] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Any modifications, equivalent substitutions, improvements, etc. made within the spirit and principles of the present invention should be included in the protection scope of the present invention.

Claims

1. The application of tung oil meal in reducing the incidence of banana wilt disease is characterized in that: The tung oil cake is used in the preparation of a banana growth regulator for inhibiting banana wilt disease; the banana wilt disease is caused by banana wilt disease physiological race 4 Fusarium oxysporum f.sp. cubense 4.

2. The use according to claim 1, characterized in that: The tung oil cake increases the activities of superoxide dismutase SOD and polyphenol oxidase PPO in banana roots to reduce the incidence of banana wilt disease.

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