112 results about "Zoospore" patented technology

The invention discloses a method for generating sporangia and releasing zoospore by inducing phytophthora capsici. The method includes the steps that phytophthora capsici is activated, an inducing solution for generating sporangia is prepared, sporangia are induced to be generated, and zoospore is induced to be released. The inducing solution used in the method is originally imported V8 vegetable juice with the volume concentration being 10-15% and a CaCO3 solution with the volume concentration being 0.2-0.3 g/L. When phytophthora capsici is induced to generate the sporangia, continuous light culture is carried out under a white fluorescent lamp. The method is simple, convenient to use, quick, free of pollution and capable of being used in phytophthora capsici pathogenicity measurement and biological activity measurement, achieved through an antibacterial agent, of phytophthora capsici.

The invention provides a method for constructing a manually controlled submarine algae field. The method comprises the steps of selection and treatment of seaweed substrates, culture of seedlings of seaweeds, sowing and the like, wherein the seaweed substrates are breeding screens formed by weaving reddish brown ropes; the step of culture of seedlings of seaweeds comprises the substeps of collection of seedlings of seaweeds and management of culture of seedlings; after the water density of the collected spores during release in the substep of collection of seedlings of seaweeds reaches 10-15 zoospores in the field of 100-times microscope, the seaweed substrates are put in the seawater to adhere; and the adhesion density is 20-30 embryo spores in the field of 100-times microscope. The method has the following advantages and beneficial effects: the variety, density, place and sowing time of the submarine algae field achieve manual control; the fixed substances of the algae, such as C, N, P and the like, are taken from the sea to the land by harvesting the seaweed, so the effect on solving the ecological problems is longer; the cost is far lower than that of the artificial reefs; thenatural submarine geomorphology is not damaged; and the submarine algae field can be formed in a shorter time, thus rapidly increasing the resource quantity.

The invention relates to a single spore isolation method of phytophthora capsici zoospores. The single spore isolation method comprises the steps that phytophthora capsici is inoculated to a V8 culture medium, sealing is performed with a sealing membrane, culture at the constant temperature of 25 DEG C is performed for 5 days, then the sealing membrane is removed, irradiation is performed for 24 hours, sterile water of 4 DEG C is added to perform induction and constant-temperature release respectively for 25 minutes so as to obtain a zoospore suspension, the suspension is diluted to reach the standard that 1-2 spores exist in each (10*) view, 1 mL of the suspension is taken and added to a V8 culture medium flat plate containing antibiotics in an evenly coated mode, constant-temperature standing is performed for 6 hours, a single germinated zoospore is found through an inverted microscope and is cut by using a scalpel to form a 0.5 cm square mark, it is confirmed that single spores are transferred to a flat antibiotic plate, and purified spores are single spore plants. The single spore isolation method is simple and convenient to operate, high in accuracy and good in rapid separating effect, adopts simple devices and can separate single spores of a large number of phytophthora capsici zoospores within a short time.

The invention belongs to the technical field of plant protection, and discloses a method of inducing phytophthora nicotianae breda de haan bacteria to generate sporangiums and release zoospores. The method comprises the following steps: activating the phytophthora nicotianae breda de haan bacteria, preparing the induction liquid generating sporangiums and inducing to generate sporangiums and release zoospores. According to the invention, the used induction liquid is the Bache wild vegetable composite fruit and vegetable juice with the volume concentration of 15-25 percent and the compound fertilizer solution with the mass concentration of 0.05-0.20 percent, and when the phytophthora nicotianae breda de haan is induced to generate the sporangiums, the continuous illumination culture is performed under the white fluorescent lamp. The method provided by the invention is simple, convenient, rapid and pollution-free, and can be used for the researches on virulence determination of the phytophthora nicotianae breda de haan bacteria, the biological activity assay of the phytophthora nicotianae breda de haan bacteria by the sterilizing agent, the evaluation of disease resistance of an epidemic disease by the luffa varieties and the like.

The invention discloses a method for inducing phytophthora parasitica var. nicotianae to generate zoosporangia and release spores. The method comprises steps of: culturing phytophthora parasitica var. nicotianae mycelia on an oat medium prepared in a narrow mouth conical flask; preparing an induction liquid for generation of zoosporangia by selecting roots of tobacco seedlings, with 6-7 pieces of true leaves completely expanding, of a black shank infected variety; inducing the mycelia with the induction liquid to generate zoosporangia and to release spores. According to the invention, a characteristic that a black shank infected tobacco variety is more easily to generate affinity reaction with phytophthora parasitica var. nicotianae is employed, and a filtrate of seedling roots of the black shank infected tobacco variety is used as the induction liquid, so as to induce the cultured mycelia to generate zoosporangia. Proved by tests, the invention has a short induction time, high zoospore release rate, high obtained zoospore content; and scraping mycelia for culture is not needed. Therefore, the invention is of less pollution and easy to operate.

The invention relates to a molecule detection technique of cayenne pepper phytophthora capsici polygalacturonase (Pcipg) 5 genes, belonging to the field of molecular biology. A pair of specific primers are designed and synthesized by the cayenne (sweet) pepper phytophthora capsici polygalacturonase (Pcipg) 5 genes, the specificity of a plurality of tested bacteria species is verified by PCR, and then the sensitivity of the specific primers is verified by carrying out quantitative molecule detection for a soil cayenne pepper phytophthora oospore and a zoospore; and on the basis, the qualitativeand quantitative molecule detection and the early warning of cayenne pepper phytophthora capsici contained in cayenne pepper diseased field soil, diseased stems, diseased leaves and diseased fruits are carried out. The invention can detect and early warn the cayenne pepper phytophthora capsici in different periods and pathopoiesia characters thereof, thereby being convenient to establish a comprehensive prevention and control technology strategy, determining and selecting an optimal prevention and control period to carry out comprehensive prevention and control and high-efficiency managementfor the cayenne pepper phytophthora capsici, really controlling the infection and the harm of pathogenic bacteria from a source and enhancing the comprehensive prevention and control effect of the cayenne (sweet) pepper phytophthora capsici.

The invention provides a primer for detecting the soybean phytophthora, and a kit and a method thereof. The primer and the detecting method for detecting the soybean phytophthora have high accuracy, high specificity and high sensitivity, and can detect the various propagules of the soybean phytophthora such as hypha, oospore, zoospore and the like. The whole technique is fast, sensitive and accurate, which has important significance in the prevention of foreign soybean phytophthora, the detection of pathogenic bacteria in domestic affected area, the international trade and the like.

The invention discloses a method for phytophthora nicotianae inoculation of tobacco. The method for the phytophthora nicotianae inoculation of tobacco includes steps that (1) preparing for tobacco seed plantlets; (2) preparing phytophthora nicotianae zoospore suspension; (3) irrigating the roots to inoculate; (4) counting the morbidity situation, and evaluating the disease resistant result. The zoospore inoculated to the roots of the seed plantlets can be quantified rigidly, the resistant difference between the same variety due to excessive inoculation amount is avoided, and the resistance identification is more precise and effective; in the actual operation, the inoculation can be performed in quantities, the space usage is small, the labor cost is saved, and the experiment period is shortened. The method for the phytophthora nicotianae inoculation of tobacco is suitable for popularization.

The invention discloses a high-efficiency biocontrol strain screening method taking pepper phytophthora blight as a target. The high-efficiency biocontrol strain screening method comprises the following steps: putting 30g of ceramsite in a tissue culture vessel, adding 40ml of plant nutrient solution to be right over the ceramsite, planting germinated pepper seeds, subjected to biocontrol seed soaking treatment, on the ceramsite, and inoculating 5ml of zoospore suspension liquid with the adjustment concentration of 105mL<-1> below the level of the nutrient solution after the pepper grows to have four main leaves. The zoospore suspension liquid is obtained by carrying out low-temperature induction after intermittently irradiating phytophthora capsici on a CA culture medium for 6-8 days. The application range of the high-efficiency biocontrol strain screening method includes screening of biocontrol strain taking pepper phytophthora blight as a target, and the like. The method is simple to operate and short in the period from seedling to inoculation, the zoospore inoculation can be conveniently implemented and observed, and the method has practical guidance significance in the biological control research work of plant diseases.

The invention disclosed a method for collecting and cultivating offspring seed of wild monostroma nitidum. The method comprises the following steps: building an appropriate seed bed and adopting an appropriate seed net to ensure that zoospores of offspring seed of wild monostroma nitidum are attached to ropes of the seed net for growing into thallus, so as to build appropriate living and growing space for wild monostroma nitidum; then managing the seed bed to ensure high yield of the wild monostroma nitidum on the seed bed and high content of chlorophyll of the wild monostroma nitidum and less sediment containing of gained wild monostroma nitidum, thereby improving the quality of the wild monostroma nitidum; the method solves the defects of long seed raising period in a man-made chamber, slow growth, higher input cost, high management difficulty and the like; the method can be applied to large scale production, the seed bed can be repeatedly used, the production cost is low, the thallus on the seed net can be easily collected to reduce the waste of mudflat and reef collection; in addition, the obtained monostroma nitidum is natural wild monostroma nitidum and is ideal green food.

The invention particularly discloses a culture medium for prompting Phytophthora capsici Leonian to generate zoosporangium, belonging to the technical field of culture mediums. The culture medium is prepared from the steps of slicing 200g of peeled carrots into pieces, boiling in 1500ml of boiling water for 30 minutes, filtering, extracting filtrate, adding water into the filtrate until the solution is 1000ml, adding 15g of agar, uniformly stirring, carrying out sterilization at 121 DEG C for 15 minutes, so as to obtain the culture medium for prompting Phytophthora capsici Leonian to generate zoosporangium. The culture medium is prepared from yellow carrots and is capable of effectively inducing and prompting the Phytophthora capsici Leonian to generate zoosporangium, so that the problem that zoosporangium is very difficulty generated from Phytophthora capsici Leonian strains P2 and P3 on a red carrot culture medium is solved, and the inoculation requirement of a lots of zoospores can be effectively met, thereby being favorable to the quantitative research of researchers on pathogenicity of these strains.

The invention provides a PCR (Polymerase Chain Reaction) detection primer for phytophthora capsici leonian. The primer comprises Enl4s and Enl1a (as shown in Seq ID No.1 and 2). Four pairs of specific primers capable of quickly detecting phytophthora capsici leonian germs can be obtained based on the sequence difference between the Enl and Ypt genes of the phytophthora capsici leonian and other phytophthora nicotianaes: Enl2s/Enl3a, Enl4s/Enl1a, Ypt2s/Ypt3a and Ypt2s/Ypt5a, wherein the sensitivity of Enl4s/Enl1a is the highest. A PCR detection system is established on the basis, which is capable of quickly and accurately detecting the phytophthora capsici leonian from complex pathogenic bacteria environments in the tissues of plants having diseases and soil. A detection kit established according to the method is simple and convenient for operation, good in specificity and high in sensitivity; and the detection kit is capable of detecting the propagules in various forms of the phytophthora capsici leonian, such as hypha, oospore, zoospore and the like, thereby having great significance in the aspects of early warning on the epidemic situation of the phytophthora capsici leonian, pathogen supervision on the epidemic area and the like.

The invention designs a molecular detection primer special for phytophthora sojae, which adopts R-SNARE protein coding gene Ykt6 needed by cell survival as a target. The primer has high sensitivity and is matched with a universal primer Ykt6F/Ykt6R of Ykt6 phytophthora; a nested PCR method is adopted, so that the sensitivity can be improved by 1000 times and trace phytophthora sojae can be detected; according to the method, specificity and sensitivity are high and hypha, oospore and zoospore of the phytophthora sojae can be detected. Simultaneously, the special for the phytophthora sojae is utilized for developing the detection method for real-time quantitative PCR and a linear regression equation, the amount of infected phytophthora sojae in soybean can be accurately and quantitatively detected and the content of the oospore of the residual phytophthora sojae in the soil can be detected so as to provide forceful data for prediction and forecast of diseases and provide technical support for entry and exit quarantine.

The invention discloses a method for Phytophthora sojae to inoculate sojae. The method for Phytophthora sojae to inoculate sojae provided by the invention is a method in which the spore suspension of the Phytophthora sojae is used to inoculate a sojae seedling of which the radicel exposes out of the seed capsule. A swarm spore root inoculating method of the invention can do inoculation strictly and quantificationally, and avoids overlarge inoculum concentration to result in that the original disease resistant variety is in susceptibility, thereby ensuring to be more effective and accurate in the application of the filtration of the anti-source; at the same time, the method of the invention has the advantages of small occupied space, less labors, short experiment period and simple latter material treatment in the actual operation, therefore, the method of the invention is suitable for popularization and application.

The invention discloses a real-time observation method for phytophthora nicotianae infection process. The real-time observation method for the phytophthora nicotianae infection process includes that preparing for tobacco seed plantlet, preparing phytophthora nicotianae zoospore suspension, irrigating the root to inoculate, dyeing and observing. According to the real-time observation method for the phytophthora nicotianae infection process, phytophthora nicotianae inoculation can be performed in a manual climatic box or a culture room by means of culture dish sponge and filter paper for preserving moisture, and the dyed and discolored seed plantlet radicle can be directly observed under a microscope. The real-time observation method for the phytophthora nicotianae infection process is convenient and easy to operate and suitable for observing the phytophthora nicotianae infection process of tobacco and detecting the resistance of different varieties.

The invention belongs to the field of microorganisms, and particularly relates to a method for separating monosporangiums of potato late blight pathogens. On the basis of an existing suspension liquid separating method, the monosporangiums are separated through the characteristic that each monosporangium contains a plurality of zoospores, and genetic materials of the zoospores are the same; the monosporangiums are stimulated at the low temperature to release the zoospores, and the new method for separating the monosporangiums is formed. The success rate of the improved method is nearly 6 times that of the original method, and operation time is halved; meanwhile, operative difficulty is greatly reduced, and the method is an effective method for rapidly separating a large mass of monosporangiums in a pathogeny-group researching test.

The invention provides a method for cultivating undaria pinnatifida and using the undaria pinnatifida to treat red tide. The method is characterized in that according to a sign before the occurrence of the red tides, that is, the temperature is increased by 2 DEG C within 7 days, a method different from a biological restraint method is adopted; before the outbreak of the red tides, the cultivatedundaria pinnatifida is added into the sea to absorb nutritional components in the sea water to alleviate the eutrophication of the sea water, and thus the occurrence of the red tides is avoided. The method includes: performing induced cultivation on female gametophyte to obtain young sporophyte, and carrying out aerated feeding cultivation to obtain mature sporophyte; releasing the sporophyll of the sporophyte into the sea to obtain zoospores, and placing seedling curtains attached with the zoospores in fresh sea water; when the germinated zoospores develop into female gametophyte, co-cultivating male gametophyte and the female gametophyte attached to the seedling curtains to complete the development and fertilization process; transferring the young sporophyte to a relevant sea area for cultivation after the young sporophyte reaches 50-100 micrometers to obtain the undaria pinnatifida and treat the red tides.

The present invention provides a phytophthora culture medium and a preparation method thereof, wherein 20-40 g of rice is added to 400-600 ml of water, water boiling bath heating is performed for 10-20 min to obtain rice water, the rice water is mixed with 400-600 ml of a soil leaching solution and 4-6 g of sunflower seed powder, and sterilization is performed to obtain the finished product. According to the present invention, the phytophthora culture medium provides a good culture effect for phytophthora, and the number of the zoosporangium of the phytophthora is more; and the phytophthora culture medium preparation method has characteristics of simpleness, wide raw material source, and simple and easily-available raw material, and is suitable for industrial production preparation.

The invention discloses a method for evaluating the pepper phytophthora blight resistance of a crop. The method comprises the specific steps of preparation of seedlings, preparation of phytophthora capsici zoospore suspension liquid, root-irrigation and inoculation, statistics of morbidity and evaluation of a disease-resistant result. Quantitative phytophthora capsici zoospores are inoculated into the seedling root parts of the crop, it is avoided that the same variety anti-infection shows difference due to excessive inoculation amount, and during resistance identification, the method is more accurate and effective; in the actual operation, inoculation can be conducted in a large batch mode, and the method has the advantages of being small in occupied space, saving labor cost and shortening test period. Accordingly, the method is suitable for application and popularization.

The invention relates to the technical field of biology, in particular to a downy mildew bacterium culture and observation method. The method comprises the following steps: collecting downy mildew bacteria; preparing sporangium suspension liquid; and culturing the sporangium suspensions. Through repeated tests, a technology capable of inducing sporangium of cucumber downy mildew to germinate andrelease zoospore is obtained, the zoospore forms static spores through movement and morphological changes, the static spores germinate to form appressorium, and the appressorium germinates to generateinfection filaments. The technology can provide an effective technique for researchers to deeply research downy mildew biological basis, disease resistance identification, bactericide screening and the like. By means of the method, researchers can achieve quantitative statistics and monitoring of the number of zoospore of downy mildew, morphological changes and the number of formed infectors, andresearch of the action mechanism of the bactericide can also be achieved. Several development stages which must be experienced before downy mildew bacteria invade a host are clearly observed. The systematic observation method can provide first-hand data for prevention and treatment of the pathogenic bacteria on symptomatic drugs.