265 results about "Pinewood nematode" patented technology

The invention discloses a technology for artificial domestication and breeding of sclerodermus and applied to prevention and treatment of pine sawyer beetles. The technology for artificial domestication and breeding of sclerodermus comprises the following steps of: firstly, feeding the sclerodermus to be substituted for the host of a barley pest; secondly, carrying out the selecting and refrigerating treatment of female sclerodermus; thirdly, packing the initiated pupa barley pest and the female sclerodermus into a finger-shaped tube, and plugging the tube with an ordered pine plug; and finally, neatly stacking and placing the tubes into a weather cultivation box with a cultivation temperature and conditions as well as prompts. In a bursaphelench xylophilus epidemic area, the throwing technology of sclerodermus comprises the following steps of: firstly, setting bait woods to induce the pine sawyer beetles to lay eggs; then examining the parasitic situation of the pine sawyer beetles in a pine tree section in an appropriate weather condition, and releasing the sclerodermus according to the groove number of the sawyer beetles; and finally investigating the parasitic situation of the sclerodermus. By the adoption of sawyer pests to prevent and treat the pine sawyer beetles so as to inhibit the occurrence of a bursaphelench xylophilus disease, the environment can not be polluted, the ecological balance is maintained, the biological diversity is kept, and the technology is an advanced technology for controlling the pine sawyer beetles and the bursaphelench xylophilus disease.

The invention provides a Monochamus alternatus adult attractant. The Monochamus alternatus adult attractant comprises an artificially-synthesized compound, and the compound is prepared through adding reagents comprising undecyl alcohol, thionyl chloride, glycol, metallic sodium and the like. The Monochamus alternatus adult attractant comprises the artificially-synthesized compound, so the effective application amount is less; the Monochamus alternatus adult attractant has a high trapping effect on Monochamus alternatus, and can be used as an effective means for monitoring and controlling the Monochamus alternatus to inhibit and delay the propagation of the pine wilt disease; and the Monochamus alternatus adult attractant generally has the advantages of simple components, low production cost, safety, environmental protection, high insect trapping efficiency, and advantage obviousness.

Two pairs of primer are designed: Pl/P3 is an idiocratic primer for pine wood nematode; P2/P3 an idiocratic primer for quasi pine wood nematode. The kit includes 1.25ml solution I,1ml solution II, 100U tag enzyme, and 50ul for each contrapositive DNA of wireworm. The solution I is as following constitutes: Worm Lysis Buffer, 500mM KCl, 100mMTris-ClpH 8.0, 15mM MgCl2, 10mMDTT, 4.5% Tween20, 0.1%gelatin, 0.2ug/ul protease K. The solution II is as following constitutes: 2xPCR reaction Buffer, 4.0mM MgCl2, 0.2mM dNTP, 0.8 mu MP1, P2, P3. The invented kit possesses high specificity, sensitiveness and stability, providing fast, sensitive and specific technical method.

The invention discloses a method for preparing bacteria-free pine wood nematodes. The method for preparing the bacteria-free pine wood nematodes comprises the following steps: culturing the pine wood nematodes; preparing pure pine wood nematode eggs; soaking the pine wood nematode eggs in 15% H2O2 for 50-60 minutes; incubating the pine wood nematodes; and culturing the pine wood nematodes and performing bacteria-free identification, thereby preparing the bacteria-free pine wood nematodes. As for the method for preparing the bacteria-free pine wood nematodes, the property of being capable of being adhered on a glass vessel of the pine wood nematode eggs is utilized, a small culture dish is adopted for collecting the eggs, the newly prepared 15% H2O2 is further used for performing sterilization treatment, the bacteria-free pine wood nematode eggs are obtained, the bacteria-free eggs are incubated, mass propagation is further performed on botrytis cinerea, and then the bacteria-free pine wood nematodes are finally obtained. Under sterile conditions, the obtained bacteria-free pine wood nematodes are used for performing an inoculation test on seedlings cultured by spruce bacteria-free tissues, and the result shows that the inoculation of the bacteria-free pine wood nematodes can still enable the bacterial-free seedlings to wilt. The method can be adopted for enabling the pine wood nematodes to be bacteria-free and lose pathogenicity, thereby being the simple and successful method.

The invention discloses an injection containing emamectin benzoate and application of the injection. The injection is prepared from the emamectin benzoate, chitosan oligosaccharide, an organic solventand a surface active agent, wherein the weight ratio of the emamectin benzoate to the chitosan oligosaccharide is 20:(1-100); the emamectin benzoate accounts for 0.1wt%-10wt% of the injection; the deacetylation degree of the chitosan oligosaccharide is equal to or higher than 50%; the organic solvent is lower alcohol which contains 1 to 8 carbon atom chains; the organic solvent accounts for 0.1wt% to 95wt% of the injection. The injection is suitable for trunk injection, various pests of woods are prevented, and the injection has the advantages of being low in cost, high in prevention efficiency and wide is application range. The injection is suitable for prevention and control over pests and diseases of the woods, and especially for prevention and control over monochamus alternatus and other wood borers of pine trees, and prevention and control over pine wood nematode, and therefore, the pine wood nematode disease is controlled and prevented.

An aseptic culture method for pine nematode includes such steps as culturing Pseudomonas fluorescens GcM5-1A (CCTCC No.M204065), deactivating, culturing the calli of black pine, deactivating, preparing the aseptic ova of pine nematode, preparing the culture liquid from deactivated cells and calli and aseptic water, adding said aseptic ova, and culturing at 15-40 deg.C for 4-20 days.

A process for earlierly detecting pine nematosis includes inoculating the disc fungus (Ps0202) or other microbial fungi on culture medium, drilling circular hole on the pine trunk to be detected, inserting the test tube containing said fungi in said hole, staying it in the hole for 3-5 days, and determining by microscope. Its advantages are easy operation and high sensitivity.

The invention provides a pine wilt disease tree identification and positioning method and system, and the method comprises the steps of obtaining the suspected disease source points of pine wood nematode according to a visible light image of a to-be-identified region; obtaining a visible light stereo image corresponding to the suspected disease source point, and obtaining a confidence coefficientof the suspected disease source point according to the visible light stereo image; judging whether the suspected disease source point is a disease source point or not by combining the visible light time sequence image, the multispectral image or the transpiration rate image corresponding to the suspected disease source point according to the confidence coefficient of the suspected disease source point; if it is judged that the suspected disease source point is a disease source point, obtaining the position information of a disease tree where the disease source point is located, and generatinga first remote sensing image with the position information of all the disease source points and corresponding pine wood nematode disease information in a to-be-identified area; and sending the first remote sensing image to an auditor for manual audit. According to the method, multiple images are utilized in the process of confirming the disease source points, the suspected disease source points are verified for multiple times, the recognition rate of the disease source points is high, and the positioning precision is high.

The present invention relates to a process for preparing and application of bionematocide and belongs to technique domain of biological pesticide. Product of the invention can be gained from conventional method that the produced strain is cultivated in test tube, enlarged cultivated in liquid and extracts active constituents of nematocide, whose produced strain is Lecanicillium psalliotae YMF1.00112, with conservation number CGMCC No.1312. The formulation of enlarged liquid base in the invention is, dextrose 10g, ammonia sulfate 1g, magnesium sulfate 1g, ferrous sulfate 0.015g, leachate of potato 1000mg, pH6.5. the invention can poison pine wood nematode and Panagrellus redivivus and be used as bio-prevention to plant parasitic nematode. The product has the advantage of high poison effect, safety to human body and environment protection, etc.

The invention discloses a method by the adoption of biological control. According to the method, the populations of the monochamus alternatus of pine forests are obviously reduced and controlled, and the number of pines which die from bursaphelenchus xylophilus is also obviously reduced. Drone pupae of bombus hypocrita are adopted as hosts of sclerodermus sp., meanwhile proliferation conditions are regulated and controlled, and therefore the parasitic ability of the sclerodermus sp. obtained through proliferation is remarkably improved.

The invention discloses a pine wood nematode detecting reagent box which takes 5'ACC ATT CGG TTG GCT CTG TT 3' as a forward primer (F), takes a 5' CCC TAA GGC GTC GGT GAA C 3' as a reverse primer (R) and takes 5' FAM- TAG CTG AGC ATC TTT T-TAMRA 3' as a fluorescent probe (P). The invention also discloses a method for adopting the reagent box to detect the pine wood nematode. The primers and the probe in the pine wood nematode detecting reagent box of the invention have higher specificity, sensitivity as well as stability and provides a fast, sensitive and specific method for detecting the pine wood nematode.

A method for quickly detecting pathogenic threadworm of pine wilt disease includes designing each pair of specific primer for PCR detection of pine threadworm and pine imitate-threadworm, obtaining single threadworm DNA as template by mechanical grinding and filtering paper absorption, carrying out specific PCR amplification of pine threadworm and pine imitate-threadworm as well as detecting PCR product.

The present invention discloses a primer and a method for detecting a specificity of a pine wood nematode in a Chinese red pine died wood of an illness, and an application thereof, wherein a nucleotide sequence of an upstream primer of the specificity detection primer is represented by SEQID NO:1, a nucleotide sequence of a downstream primer is represented as SEQ ID NO:2. the primer of the invention is capable of separating the pine wood nematode from other 12 nematodes of the Chinese red pine fat wood and Chinese red pine as well as common four fungus and ten bacteria of the Chinese red pine. the invention also provides a method for implementing a PCR amplification of the pine wood nematode of the diseased wood directly without separation of the pine wood nematode from the diseased wood, after rubbing the diseased wood liquid nitrogen, by means of two cleavages of WLB and protease K, a DNA formwork for the PCR amplification will be obtained after purifying, the amplification efficiency will achieve 100%, a separation time is saved and a detection efficiency is improved. The detection method of the invention is simple and high-efficiency, has a high accuracy and the specificity; it is capable of being produced into reagent kits to popularize widely.

The invention relates to nematocide and its appliances thereof, belonging to the field of biological pesticide technique. Said nematocide is acquired through the sequence of culturing preparing strain in test tube, fluid enlargement culture and extracting nematocide active component, and said preparing strain is as follows: germinated barley-acceptable single cell bacteria stenotrophomonas maltophilia G6 which is preserved in microbe bacterial preservation management commission common microbe center of China with preservation data: 2005-10-31 and preservation registration number CGMCC No: 1522. The main mechanism of action of G6 strain is to break up body wall of worm through producing enzyme and poisoning worm, especially pine beam worm, and could be used to prepare biological nematocide. The production in said invention is characterized on high poison ability, which has better potential utility.

The invention is suitable for the field of nematode disease control and provides a method for controlling pine wood nematode diseases by utilizing body surface bacteria of American pine wood nematodes. The method comprises the steps of: spraying American bacterium suspension liquid into woodlands: spraying once in May for two years in a row, wherein the body surface pathogenic bacteria of the pine wood nematodes are replaced with American bacteria in two or three years so as to control the prevalence of woodland pine wood nematode diseases in China. According to the invention, the biological control is carried out by utilizing the replacement of ecological niche of the body surface bacteria of the pine wood nematodes and bacterial strains isolated and screened from body walls of the American pine wood nematodes. In the method, the biocontrol bacteria sprayed into the woodlands have the characteristics of low pathogenicity, strong reproduction and replacement capabilities and the like and are capable of rapidly replacing the body surface strong pathogenic bacteria of the pine wood nematodes in China to ensure that the pine wood nematodes lose the pathogenic ability and the purpose of controlling the prevalence of pine wood nematode diseases is achieved. The method has the advantages of simpleness, practicability, easiness in popularization and application, lower cost, good and thorough effect and the like.

According to the technical scheme, the sky-ground integrated monitoring method comprises the steps that S1, firstly, infected and withered trees are extracted through unmanned aerial vehicle images, plaques of the infected and withered trees are extracted through satellite images, and on-site marking, positioning and measuring are conducted on the withered trees through ground investigation; s2, respectively acquiring high-spatial-resolution satellite remote sensing data and unmanned aerial vehicle remote sensing data as data sources, and jointly forming a data set of a monitoring area in combination with ground survey data and forest resource second-class survey data; s3, establishing samples of the epidemic trees according to the data, wherein the samples comprise spectral features, texture features and geometric features of the pine wood nematode disease withered and dead trees; and S4, according to the sample and satellite remote sensing, confirming the infected, withered and dead tree plaques, comprehensively applying an image enhancement and image classification method, performing verification, and establishing spatial distribution information and position information of the pine wood nematode disease withered and dead trees. The method has the advantages of high epidemic area screening efficiency, capability of realizing large-area efficient monitoring, convenience in acquisition of remote sensing data of the unmanned aerial vehicle, high recognition precision and guarantee for precision verification.

The present invention relates to a method for controlling pine wood nematode by installing, at regular intervals, net type traps including an attractant composition for attracting Monochamus alternatus and Monochamus saltuarius, which are pollinating insects of pine wood nematode, and capturing, within the traps, the attracted Monochamus alternatus and Monochamus saltuarius. The present invention can constantly and effectively capture Monochamus alternatus and Monochamus saltuarius over a whole generation by forming a net-type trap system at a control area on the basis of the physiological, ecological, and behavioral characteristics of Monochamus alternatus and Monochamus saltuarius which do not show active scattering mobility, and thus can control the damage caused by pine wood nematode through the medium of Monochamus alternatus and Monochamus saltuarius and can forecast whether or not damage will occur.

The invention relates to the technical field of insect pest prevention and treatment in forestry, in particular to a method for comprehensively preventing and treating longhorn beetles. In the method,first, healthy masson pine wood is selected, pine wood nematodes are introduced and utilized for infecting the masson pine, then the infected masson pine and beauveria bassiana are blended and subjected to anaerobic digestion, yeast is obtained, monoterpene pheromones of the pine wood nematodes are carried in the masson pine yeast after infection of the pine wood nematodes, the pine wood nematodes are hosts of the longhorn beetles, the pheromones of the hosts will lure and gather the longhorn beetles, the lured and gathered longhorn beetles will make contact with beauveria bassiana, in this way, the longhorn beetles and larvae of the longhorn beetles can be effectively killed, the longhorn beetles will also carry the deadly beauveria bassiana to other places to promote diffusion of the longhorn beetle deadly bacteria, and then the killing rate is further increased for the longhorn beetles. Sclerodermus harmandi as parasitical natural enemies of the longhorn beetles is released aroundluring wood filled with self-made luring and killing agents and is parasitic on the larvae of the longhorn beetles to further increase the killing rate for the longhorn beetles, finally, the luring wood is burned down in a concentrated mode, the longhorn beetles are thoroughly killed, and broad application prospects are achieved.

The pesticide for killing pine material nematode consists of abamectin 0.001-0.1 wt%, copper sulfate 30-35 wt%, Shenzhanling 1-3 wt% and water the rest. Of the components, copper sulfate can kill separated nematode directly in high effect, wide killing spectrum and lasting effect, Shenzhanling makes copper sulfate permeate into pine material to kill nematode, and abamectin can kill nematode and black pine longicorn. The present invention has the advantages of high permeation, high efficiency, last effect, safety and convenience.

The invention provides an attractant for entrapping imagoes of arhopalus rusticus, the attractant comprises the components including 3-carene and absolute ethyl alcohol in a volume ratio of 0.8-1.2:1. The attractant has advantages of simple preparation technique, obvious entrapping effect on arhopalus rusticus, no pollution, environmental friendliness, good entrapping persistence and the like; the attractant is used together with polyethylene thin-walled slowly-releasing plastic bottles and black plastic cross traps, and has an excellent entrapping effect on imagoes of arhopalus rusticus; theattractant is capable of predicting, forecasting, preventing and treating arhopalus rusticus, as well as effectively controlling the density of arhopalus rusticus in forest and preventing the spread and expansion of nematode disease for pine; therefore, the attractant provides an important technical and product support for the comprehensive treatment of pine wood ematode disease, pine wood mucronatus and arhopalus rusticus.

The invention discloses a kit for qualitative detection of pinewood nematode and a detection method thereof. Reagents of the kit comprise a DNA extract, a reaction liquid of constant-temperature amplification of pinewood nematode nucleic acid, a positive control and a negative control, wherein the positive control is a plasmid containing a DNA fragment of a pinewood nematode BxS1 gene, and the negative control is sterile double distilled water. The kit detects the pinewood nematode through constant-temperature amplification of nucleic acids, and sensitivity thereof is that 100 copies can be detected in each reaction system. Repetitive experiments show that detection results have no significant differences and are good in repeatability. Only 2 hours are required to finish the sample detection, thereby greatly reducing detection time. Meanwhile, only one person is needed to finish all operation process of the kit, and one to hundreds of samples can be detected for one time, thereby reducing manpower waste, meeting requirements of rapid detection of the pinewood nematode and thus being quite practical.

The invention relates to the use of alpha-terthiophene, in particular the use for the control of pine wilt disease, experimental investigation has shown that, when the when the concentration of the alpha-terthiophene is higher than 10-4g/L, the pine wilts can be poisoned within short period of time. The alpha-terthiophene can be applied to pine trees directly or with the compositions containing the alpha-terthiophene.