Application of camphene in inducing resistance of Panax notoginseng and alleviating leaf black spot disease

Abstract

The invention provides an application of camphene in inducing resistance of Panax notoginseng and alleviating leaf black spot disease, and relates to the technical field of Panax notoginseng planting. Camphene is placed on continuously planted soil to fumigate Panax notoginseng in advance, so as to induce resistance of Panax notoginseng to leaf black spot disease infection in the later stage. It can be seen from the test results that when Panax notoginseng induced by camphene fumigation is infected by black spot pathogenic bacteria again, the resistance of Panax notoginseng can be significantly enhanced and the incidence area of ​​black spot disease can be reduced.

Description

Technical Field

[0001] The invention relates to the technical field of notoginseng planting, and particularly to application of camphene in inducing notoginseng resistance and alleviating leaf black spot disease. Background Art

[0002] Panax notoginseng is a perennial herbaceous plant of the genus Panax, family Araliaceae, order Apiaceae. It is also known as ginseng, field notoginseng, field notoginseng, mountain lacquer, panlongqi, golden notoginseng, and copper skin and iron bones. It is recorded in ancient medical books such as "Compendium of Materia Medica", "Bencao Qiuzhen", "Bencao Congxin" and "Compendium of Materia Medica Supplement". It has the characteristics of warm nature, sweet and slightly bitter taste, and has good effects such as removing blood stasis and stopping bleeding, promoting blood circulation and relieving pain. It is a good medicine widely recognized by the medical community. It is mainly used as medicine in the form of dried tubers and flowers, and there are various ways to process and eat it. Due to its high medicinal value, wild picking resources can no longer meet the growing social demand, so modern artificial farmland cultivation is widely adopted.

[0003] So far, the development of the Panax notoginseng planting industry has made great progress, but there is a big gap compared with traditional crop planting, especially the growth problems of Panax notoginseng caused by continuous cropping obstacles have seriously restricted the sustainable development of the Panax notoginseng industry. In production practice, the occurrence of continuous cropping obstacles usually leads to problems such as reduced germination rate of Panax notoginseng seeds and limited growth of Panax notoginseng seedlings. Black spot disease is also a problem encountered in the planting process of Panax notoginseng. Panax notoginseng black spot disease is a fungal disease caused by Alternaria ginseng. The disease mainly attacks the leaves, stems, petioles, flower axes, fruits, fruit stalks, roots, rhizomes and buds of Panax notoginseng, but mainly affects the young and tender tissues of the aboveground part or the parts with loose tissue structures, accounting for more than 60% of all diseased plants. When Panax notoginseng leaves are infected with black spot disease, small, round or irregular brown spots will initially appear on the leaves. As the disease progresses, the spots will gradually expand and turn dark brown, and a dark brown mold layer may appear in the center of the spots, and the spots are easy to perforate and rupture. When the relative humidity is high, the lesions will expand rapidly and cause the leaves to fall off, which will have a significant impact on the yield and quality of Panax notoginseng. Therefore, how to induce resistance to Panax notoginseng in daily planting to reduce leaf black spot disease is not only a production problem that needs to be solved urgently, but also an ecological problem of sustainable development. Summary of the invention

[0004] In view of the shortcomings of the prior art, the present invention induces the resistance of Panax notoginseng to leaf black spot infection in the later stage by placing camphene on the continuously cropped soil to fumigate Panax notoginseng in advance.

[0005] To achieve the above objectives, the present invention is implemented through the following technical solutions:

[0006] On the one hand, the present invention proposes the use of camphene in inducing resistance of Panax notoginseng and alleviating leaf black spot disease.

[0007] Furthermore, the application method is to fumigate Panax notoginseng seedlings with camphene in a closed space.

[0008] Furthermore, the fumigation time is 1-3 days, for example, 1 day, 2 days, 3 days, preferably 3 days.

[0009] Further, the volatile concentration of camphene used for fumigation in a confined space is 0.1-20g / L;

[0010] Furthermore, the volatilization concentration of camphene used for fumigation in a confined space is 1-20 g / L, that is, the mass of camphene / volume of the confined space is 1-20 g / L, for example, it can be 1 g / L, 2 g / L, 3 g / L, 5 g / L, 10 g / L, 15 g / L, 20 g / L, preferably 5 g / L.

[0011] On the other hand, the present invention provides a fumigant for inducing resistance of Panax notoginseng and alleviating leaf black spot disease.

[0012] Furthermore, the volatilization concentration of camphene in a confined space is 1-20 g / L.

[0013] Compared with the prior art, the present invention has the following advantages:

[0014] The present invention places camphene, a volatile substance, on the continuously cropped soil to fumigate the Panax notoginseng in advance, so as to induce the resistance of the Panax notoginseng to leaf black spot infection at a later stage. Experiments show that when the Panax notoginseng induced by camphene fumigation is then infected by black spot pathogenic bacteria, the resistance of the Panax notoginseng can be significantly enhanced and the black spot area can be reduced. BRIEF DESCRIPTION OF THE DRAWINGS

[0015] In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the following briefly introduces the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying creative work.

[0016] Figure 1 The figure shows the area of ​​lesions on the leaves of Panax notoginseng after one day of fumigation induction treatment with camphene at different volatile concentrations in the embodiment of the present invention; wherein A, B, C, D, and E are the area of ​​lesions on the leaves of Panax notoginseng treated by fumigation induction treatment with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively;

[0017] Figure 2 A comparison of the areas of infection of black spot pathogens on the leaves of Panax notoginseng after one day of fumigation induction with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L in the embodiment of the present invention is shown;

[0018] Figure 3 The figure shows the area of ​​lesions on the leaves of Panax notoginseng after two days of fumigation induction treatment with camphene at different volatile concentrations in the embodiment of the present invention; wherein A, B, C, D, and E are the area of ​​lesions on the leaves of Panax notoginseng treated by fumigation induction treatment with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively;

[0019] Figure 4 A comparison of the areas of infection of black spot pathogens on the leaves of Panax notoginseng after two days of fumigation induction with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L in the embodiment of the present invention is shown;

[0020] Figure 5 The diagram shows the area of ​​lesions on Panax notoginseng leaves after three days of fumigation induction treatment with camphene at different volatile concentrations in the embodiment of the present invention; wherein A, B, C, D, and E are the area of ​​lesions on Panax notoginseng leaves treated by fumigation induction with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively;

[0021] Figure 6 A comparison of the areas of infection of black spot pathogens on the leaves of Panax notoginseng after three days of fumigation induction with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L in the embodiment of the present invention is shown;

[0022] Figure 7 The results of the disease index of reducing leaf black spot disease of Panax notoginseng by inducing Panax notoginseng with different volatile concentrations of camphene for one day, two days and three days respectively in the embodiment of the present invention are shown; wherein A, B and C are the disease index of Panax notoginseng leaf black spot disease after inducing Panax notoginseng with volatile concentrations of 0g / L, 0.1g / L, 1g / L, 5g / L and 20g / L camphene fumigation for one day, two days and three days respectively. A. panax Disease index of the infection;

[0023] Figure 8 The relative gene expression results of the key resistance genes of Panax notoginseng after three days of induction of Panax notoginseng by fumigation with different concentrations of camphene in the embodiment of the present invention are shown; wherein A, B, C, and D are the relative expression levels of four Panax notoginseng resistance genes MPK3, WRKY33, WRKY22, and Rboh D after three days of induction of Panax notoginseng by fumigation with volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively. DETAILED DESCRIPTION

[0024] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention is clearly and completely described below in combination with the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without creative work are within the scope of protection of the present invention. Embodiment 1:

[0025] Camphene-induced resistance test of Panax notoginseng:

[0026] The present embodiment adopts the confined space fumigation method to carry out the camphene induced resistance test. The Panax notoginseng seeds used in the embodiment are purchased from the Panax notoginseng trading market in Wenshan City, Yunnan Province. The seeds are collected from three-year-old Panax notoginseng, and the seed maturity is high, the grain is full, and there is no disease. After the seeds are harvested, the red skin is washed off, and the stratification moisture is moisturized to promote the seed after-ripening. After the Panax notoginseng seeds after stratification are wrapped evenly with seed coating agent, they are sown in the Daheqiao teaching practice base of Xundian, Yunnan Agricultural University (25 ° 31' 01.4 "N, 103 ° 16' 48.9 "E), and normal water and fertilizer management is maintained after emergence. After the above-ground part of the seedling is fully developed, it is transplanted in a potted plant to carry out subsequent tests, and 10 to 12 seedlings are transplanted in each pot, and the seedlings are slowed down for one week after the transplanting is completed. The monomer compound of camphene is selected from Merck Sigma-Aldrich, and its purity is greater than or equal to 95%.

[0027] The specific test process is as follows:

[0028] Select the Panax notoginseng potted plants that grow well after transplanting, remove the weeds on the soil surface, water them thoroughly and leave them for 1 day, then cover the pots with plastic bags so that only the above-ground part of the Panax notoginseng can come into contact with volatile substances, avoiding the impact of volatile substances on the soil.

[0029] The 6 treated Panax notoginseng potted plants were placed in a transparent plastic sealed culture box (the volume after deducting the pots was about 25 L, with a total of 5 boxes), and then 0g / L, 0.1g / L, 1g / L, 5g / L, and 20g / L (compound standard mass / sealed transparent box volume) of camphene monomer compound were added to the inverted centrifuge tube lid for closed fumigation of Panax notoginseng. The fumigation induction time was 1 day, 2 days, and 3 days, respectively.

[0030] After the Panax notoginseng seedlings were induced with camphene, the leaves were dipped and inoculated with black spot pathogens Alternaria panax The bacterial cake (SL15) was then moved to a transparent plastic incubator for moisturizing culture. Embodiment 2:

[0031] 1. Detection of the effect of camphene inducing Panax notoginseng to alleviate leaf black spot disease;

[0032] The diseased leaves 5 days after inoculation in Example 1 were collected, and the diseased leaves were scanned using an Epson Perfection V850 Pro scanner. The relative diseased area of ​​the induced treatment and the control treatment was then intercepted and calculated using Adobe Photoshop software to evaluate the effect of camphene inducing resistance to Panax notoginseng. IBM SPSS 19 was used to calculate the difference of the relevant data, and finally Graphpad prism9 was used for drawing and presentation.

[0033] The results of a one-day experiment using camphene fumigation to induce Panax notoginseng are as follows Figure 1 and Figure 2 As shown, Figure 1 A, B, C, D, and E are the lesion areas of Panax notoginseng leaves treated by fumigation with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively. Figure 2 The figure is a comparison of the infection area (diseased area) of the black spot pathogen on the leaves of Panax notoginseng after fumigation induction with camphene at volatile concentrations of 0g / L, 0.1g / L, 1g / L, 5g / L, and 20g / L. It can be seen that the anti-black spot effect of Panax notoginseng after fumigation induction with 0.1g / L, 1g / L, 5g / L, and 20g / L camphene is better. Compared with the treatment without fumigation induction (0g / L), the black spot area of ​​the leaves of Panax notoginseng after fumigation induction with three concentrations of camphene (1g / L, 5g / L, and 20g / L) is at most 0.67cm 2 Down to 0.33cm 2 (20g / L treatment).

[0034] The results of a two-day experiment using camphene fumigation to induce Panax notoginseng are as follows Figure 3 and Figure 4 As shown, Figure 3 A, B, C, D, and E are the lesion areas of Panax notoginseng leaves treated by fumigation with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively. Figure 4 The figure is a comparison of the infection area of ​​black spot pathogen on the leaves of Panax notoginseng after fumigation induction with camphene at volatile concentrations of 0g / L, 0.1g / L, 1g / L, 5g / L, and 20g / L. It can be seen that the anti-black spot effect of Panax notoginseng after fumigation induction with 0.1g / L, 1g / L, 5g / L, and 20g / L camphene is better. Compared with the treatment without fumigation induction (0g / L), the black spot area of ​​Panax notoginseng leaves after fumigation induction with three concentrations of camphene (1g / L, 5g / L, and 20g / L) for two days is at most from 0.64cm 2 Down to 0.22cm 2 (5g / L treatment).

[0035] The results of a three-day experiment using camphene fumigation to induce Panax notoginseng are as follows Figure 5 and Figure 6 As shown, Figure 5 A, B, C, D, and E are the lesion areas of Panax notoginseng leaves treated by fumigation with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively. Figure 6 The figure is a comparison of the infection area of ​​black spot pathogen on the leaves of Panax notoginseng after fumigation induction with camphene at volatile concentrations of 0g / L, 0.1g / L, 1g / L, 5g / L, and 20g / L. It can be seen that the anti-black spot effect of Panax notoginseng after fumigation induction with 0.1g / L, 1g / L, 5g / L, and 20g / L camphene is better. Compared with the treatment without fumigation induction (0g / L), the black spot area of ​​Panax notoginseng leaves after fumigation induction with three concentrations of camphene (1g / L, 20g / L) for three days at most increased from 0.58cm 2 Down to 0.24cm 2 (1g / L treatment).

[0036] From the above results, it can be seen that the effect of using 5g / L camphene to fumigate and induce Panax notoginseng for two days is the best.

[0037] 2. Calculate the disease index of Panax notoginseng at four volatile concentrations (0.1g / L, 1g / L, 5g / L, and 20g / L) for camphene to induce the reduction of leaf black spot disease.

[0038] The results are as follows Figure 7 As shown, A, B, and C are the black spot pathogens of Panax notoginseng leaves induced by fumigation with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L for one day, two days, and three days, respectively. A. panax Disease index of infection. Figure 7 It can be seen that the fumigation induction of Panax notoginseng with camphene at four concentrations (0.1g / L, 1g / L, 5g / L, 20g / L) for one, two or three days can improve the resistance of Panax notoginseng to leaf black spot disease and reduce the disease index. Among them, the lowest disease index for one day of camphene induction was the 0.1g / L treatment, reaching 23.6. The lowest disease index for two days of camphene induction was the 1g / L treatment, reaching 24.7. The lowest disease index for three days of camphene induction was the 5g / L treatment, reaching 22.5.

[0039] 3. Detection of resistance genes;

[0040] From the test results of 1 and 2, it can be seen that after comparison, it was found that the effect of using 5g / L camphene fumigation to induce Panax notoginseng for three days was the best. Therefore, the diseased leaves treated with three days of fumigation were selected for qRT-PCR expression measurement. The results are as follows Figure 8As shown, A, B, C, and D are the relative expression levels of four Panax notoginseng resistance genes MPK3, WRKY33, WRKY22, and Rboh D after three days of fumigation induction with camphene at volatile concentrations of 0 g / L, 0.1 g / L, 1 g / L, 5 g / L, and 20 g / L, respectively.

[0041] from Figure 8 It can be seen that among the four resistance expression genes (MPK3, WRKY33, WRKY22 and Rboh D), the 5g / L camphene fumigation-induced treatment had the strongest resistance enhancement effect and the highest relative expression level, which were significantly higher than the non-fumigation treatment (0g / L).

[0042] From the above test results, it can be seen that fumigation with camphene to induce Panax notoginseng and then infection with black spot pathogen can significantly enhance the resistance of Panax notoginseng and reduce the area of ​​black spot lesions. Among them, fumigation induction with 5g / L camphene for three days is the most preferred and has the best effect.

[0043] The above embodiments are only used to illustrate the technical solutions of the present invention, rather than to limit the same. Although the present invention has been described in detail with reference to the aforementioned embodiments, those skilled in the art should understand that the technical solutions described in the aforementioned embodiments may still be modified, or some of the technical features may be replaced by equivalents. However, these modifications or replacements do not deviate the essence of the corresponding technical solutions from the spirit and scope of the technical solutions of the embodiments of the present invention.

Claims

1. The application of camphene in inducing resistance of Panax notoginseng and alleviating leaf black spot disease is as follows: the application method is to fumigate Panax notoginseng seedlings with camphene in a closed space; the fumigation time is 1-3 days; the volatile concentration of the camphene used for fumigation in the closed space is 0.1-20g / L.

2. The use according to claim 1, characterized in that: The fumigation time is 3 days.

3. The use according to claim 1, characterized in that: The volatile concentration of camphene used for fumigation in a confined space is 1-20g / L.

4. The use according to claim 1, characterized in that: The volatile concentration of camphene used for fumigation in a confined space is 5g / L.

Images