CN104404124A - Identification method for clubroot resistance of non-heading cabbages - Google Patents
Identification method for clubroot resistance of non-heading cabbages Download PDFInfo
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- CN104404124A CN104404124A CN201410668162.2A CN201410668162A CN104404124A CN 104404124 A CN104404124 A CN 104404124A CN 201410668162 A CN201410668162 A CN 201410668162A CN 104404124 A CN104404124 A CN 104404124A
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Abstract
The invention discloses an identification method for clubroot resistance of non-heading cabbages. The identification method comprises the following steps: preparing a clubroot bacterium suspension with the concentration of 1*10<7> to 2*10<8>bacteria/ mL; when seedlings of the non-heading cabbage grow to have three leaves and one heart, uniformly irrigating the roots of the seedlings with the prepared clubroot bacterium suspension so as to soak the roots of the seedlings in the clubroot bacterium suspension for 3-4 days; taking the seedlings out of the clubroot bacterium suspension, culturing the seedlings for 40-50 days by a conventional method, and identifying and screening non-heading cabbages which are not infected by clubroot. By the identification method, when the seedlings of the non-heading cabbages grow to have three leaves and one heart, the seedlings are inoculated with clubroot bacteria by a root dipping method; by investigating the illness occurrence situation of plants, the disease-resistant or infected non-heading cabbages are identified.
Description
Technical field
The invention belongs to crop in cruciferae Disease Resistance Identification technical field, be specifically related to the authentication method of the anti-club root of a kind of Chinese cabbage.
Background technology
Crop in cruciferae club root contaminates by plasmodiophora brassica bacteria (plasmodiophora brassicae Woron) the crop in cruciferae root disease caused, this germ is a kind of obligate parasite, contaminates crop in cruciferae specially.This pathogenic bacteria invaded plants root hair, cortex, form layers, produce containing the germ secretory product of plant growth hormones, stimulates the abnormal division of parenchyma cell to increase, and expands the daetylorhiz forming warty at hyperplasia position.The appearance of daetylorhiz reduces the ability of Root Absorption and supply moisture content and nutrient on the one hand, causes over-ground part dehydration, withered and yellow and dead; Make root system of plant lose outer field provide protection on the other hand, easily invaded by other miscellaneous bacteria and cause rotten.
Club root can endanger the multiple Cruciferae cultivation such as Chinese cabbage, green vegetables, Cauliflower, wild cabbage, rape and wild plant, once occur, will cause serious harm.In China, this disease mainly occurs in the major production areas of some crop in cruciferae in East China and south China.Crop in cruciferae club root infectivity is extremely strong, can propagate with number of ways such as seed, fertilizer, rainwater, irrigation water, insect pest, the plague of rats, farming operations, pathogenic bacteria survives the winter on soil or seed, can survive in soil more than 6 years, germ all can grow at 9 ~ 23 DEG C, the most susceptible disease of soil moisture content 70% ~ 90%, this sick velocity of propagation is fast, onset area is in tens times between short several years, and even hundreds of times increase, and endanger very serious.
Chinese cabbage (Brassic campestris ssp.chinensis Makino) has another name called Plantula Brassicae chinensis, green vegetables, belong to Cruciferae Brassica genus, because its quality is tender, unique flavor, the feature such as nutritious, occupy an important position in people's lives, special in Shanghai, the middle and lower reach of Yangtze River and areas to the south, Chinese cabbage is that people must eat every day, indispensable popular vegetables.But in recent years, suburb of Shanghai, as the ground Chinese cabbage club root morbidities such as Qingpu, Songjiang, Kingsoft and Pudong increase the weight of year by year, serious region output sharply declines, and even returns product, affects the regular supply in Chinese cabbage market, causes extreme loss.Usually, a large amount of hyperplasia of root tissue exception after Chinese cabbage morbidity, expand the daetylorhiz forming warty at hyperplasia position, the appearance of daetylorhiz compromises growth and the receptivity of root system, the growth and development of plants infected is slow, makes plant wither, yellow and fallen leaves, and even whole strain is dead.To cause production declining after green vegetables are susceptible, quality reduces, and time serious, soft rotten bacterium and other pathogenic bacterias are taken advantage of a weak point, and infect the position that is injured, No kernels or seeds are gathered, as in a year of scarcity finally to make plant withered death, causes great loss to vegetable grower.
The domestic and international research to Cruciferae club root is many at present, but mainly combine in Chinese cabbage, wild cabbage, rape and radish etc., for club root Resistance Identification technique study many of Chinese cabbage, wild cabbage and rape, the Resistance Identification technique study infected after club root for Chinese cabbage is less, and the morbidity of Chinese cabbage root kind disease is on the rise, therefore be badly in need of wanting an authentication method can identifying anti-club root fast and accurately, for the raw-material screening of the anti-club root of Chinese cabbage provides technical support.
Summary of the invention
The object of the invention is, the authentication method of the anti-club root of a kind of Chinese cabbage is provided, solve in prior art the authentication method fast and accurately lacked about the anti-club root of Chinese cabbage.
The present invention is as follows for solving the problems of the technologies described above adopted technical scheme:
An authentication method for the anti-club root of Chinese cabbage, comprises the steps:
Step 1, preparation pathogen plasmodiophora suspension;
Step 2, grows to three leaves wholeheartedly periods the seedling of Chinese cabbage, the club root suspension of aforementioned preparation is carried out uniform irrigation to seedling root;
Step 3, carries out cellar culture by the seedling after aforementioned impregnation, collects the Chinese cabbage not infecting club root.
Further, in described step 1, the concentration of the pathogen plasmodiophora suspension of preparation is 1 × 10
7~ 2 × 10
8individual/mL.
Further, in described step 1, the compound method of club root suspension is: the fresh daetylorhiz collecting the Chinese cabbage of morbidity, at plant tissue crusher for crushing, filtration, the centrifugal 10min of 4000r/min after cleaning, be modulated into pathogen plasmodiophora suspension with distilled water.The fresh daetylorhiz of the Chinese cabbage of described collection and the club root suspension prepared need to preserve under-20 DEG C of conditions.
Further, after club root suspension is irrigated seedling root in described step 2, seedling root is flooded 3 ~ 4 days in club root suspension.
Further, the type of rearing of described Chinese cabbage is: the vinyl disc selecting water containing band concave-convex bottom, places the foam cave dish with multiple hole in vinyl disc, described Chinese cabbage sowing is grown in the dish of foam cave.Particularly, described vinyl disc is of a size of 45cm × 30cm × 9cm, Pan Wei77 hole, described foam cave.
Further, when the Chinese cabbage in the dish of cave grow to three leaves wholeheartedly time, the seedling root pouring 200ml club root suspension coiled in each cave; Club root suspension is flowed in vinyl disc by seedling, and preserves 3 days in vinyl disc.
Further, in described step 3, the condition of seedling cellar culture is: daytime cultivates 10 hours under 28 DEG C of conditions, and cultivate 14 hours under 18 DEG C of conditions in the evening.The time of described seedling cellar culture is 35 ~ 45 days.
Compared with prior art, beneficial effect of the present invention is as follows:
1, the present invention is by filling with root inoculation method, after the club root suspension of preparation is carried out root irrigation to Chinese cabbage seedling, disease-resistant plant root is normal, and susceptible plant root tissue paraplasm, the daetylorhiz forming warty that expands, thus disease-resistant and disease plant are made a distinction, reach the object obtaining anti-club root plant material.
2, the present invention finds that Chinese cabbage seedling wholeheartedly to club root germ susceptibility more by force, more easily infects this disease at three leaves period.Therefore, select Chinese cabbage in three leaves wholeheartedly periods, fillings root is carried out to Chinese cabbage seedling and inoculates, make the disease-resistant plant filtered out have stronger anti-infection property.
3, the present invention has not only distinguished the anti-sense ability of Chinese cabbage germ plasm resource to Cruciferae club root, and the anti-knee characteristic of disease of a certain Chinese cabbage material colony can be improved by filling with root authentication method, for the seed selection of Chinese cabbage anti-club root new variety provides more disease-resistant material, for all very valuable in the practice of Chinese cabbage breeding for disease resistance and theoretical investigation thereof.
Accompanying drawing explanation
Fig. 1 is the vinyl disc of the band concave-convex bottom being placed with 77 foam cave, hole dishes in the embodiment of the present invention;
Fig. 2 is the Chinese cabbage plant infecting club root in the embodiment of the present invention;
Fig. 3 is the Chinese cabbage plant not infecting club root in the embodiment of the present invention.
Embodiment
Below by way of specific embodiment, technical scheme of the present invention is described in detail.
Embodiment 1, Chinese cabbage infects the identification and analysis of club root at different times
To the new short green grass or young crops of the conventional variety of the Chinese cabbage of the main cultivation in Shanghai and the heat resistant variety new summer green grass or young crops carry out club root suspension root irrigation in different vegetative period (cotyledon period and three leaf one heart stages) with same time, same concentrations.
Test method: test and carry out in greenhouse, Inst. of Gardening, Academy of Agriculture of Shanghai test base.First, gather the fresh diseased tissues (i.e. the knee knurl of Chinese cabbage) of Chinese cabbage club root on Hong Yang farm, Qingpu district, be sub-packed in plastics bag after cleaning, preservation in-20 DEG C of refrigerators.Take out knee knurl during test, break into homogenate with plant tissue pulverizer, four layers of filtered through gauze, the centrifugal 10min of refrigerated centrifuge 4000r/min, abandons supernatant, and suspend with distilled water, repeat 3 times, being finally suspended into pathogen plasmodiophora concentration is 2 × 10
8individual/ml, obtained club root suspension, saves backup in 4 DEG C.
By sterile soil and matrix, (main component of this matrix is vermiculite, peat and perlite, be purchased from Shanghai Sun Qiao modern agriculture Science and Technology Ltd.) with volume ratio be 1:3 ratio mixing mix thoroughly, select the vinyl disc of water containing band concave-convex bottom, the middle foam cave dish placing 77 holes, sowing Chinese cabbage seed in the dish of cave.See Fig. 1, the figure shows the vinyl disc of the band concave-convex bottom of foam cave, placement 77 hole dish.
Grow to cotyledon period and three leaves Chinese cabbage seedling respectively and wholeheartedly carry out filling root inoculation process period.The concrete grammar of root irrigation is: measure 200ml club root suspension with graduated cylinder, evenly waters the seedling root being filled in each cave dish, and club root suspension is flowed in vinyl disc by seedling, and club root suspension preserves 3d in vinyl disc.After 3d, Chinese cabbage seedling carries out cellar culture, daytime 28 DEG C, 10 hours, evening 18 DEG C, 14 hours, after cellar culture 45d, Chinese cabbage plant is dug out, clean root, whether the incidence of investigation plant, have knee knurl according to the root of Chinese cabbage and judge whether to have infected club root.See Fig. 2, the figure shows the Chinese cabbage infecting club root.See Fig. 3, the figure shows the Chinese cabbage not infecting club root.Statistics disease plant number, calculates sickness rate.Sickness rate=morbidity strain number/total strain number × 100%.See table 1, be the result of Chinese cabbage in different growing stages infection club root of two kinds.
Table 1, Chinese cabbage infects the result of club root in different growing stages
As can be seen from Table 1: the Chinese cabbage of two kinds is only 5.3% ~ 11.2% at the sickness rate of cotyledon period club root, and three leaves wholeheartedly period, club root sickness rate can reach 57.5% ~ 86.7%.By data results, can determine that Chinese cabbage wholeheartedly to club root germ susceptibility more by force, more easily infects this disease at three leaves period.Therefore, in technical solution of the present invention, treat that Chinese cabbage seedling grows to three leaves wholeheartedly periods, carry out filling root inoculation process.
Embodiment 2, the club root Resistance Identification of the Chinese cabbage of different varieties
For examination material: all material in embodiment provides by Shanghai green vegetables seminar of academy of agricultural sciences horticulture institute.Wherein be numbered 13-1g-1 to 13-1g-52 and 13-1g-54 to 13-1g-61, be the self-mating system material of this seminar through purifying for many years; Being numbered 13-1g-65 to 13-1g-70 is intermediate materials; Numbering 13-3g-1 to 13-3g-3 is the anti-club root Chinese cabbage cultivar of the Sheng Nisi seeds company of Korea S introduced.
Test method: test and carry out in greenhouse, Inst. of Gardening, Academy of Agriculture of Shanghai test base.First, gather the fresh diseased tissues (i.e. the knee knurl of Chinese cabbage) of Chinese cabbage club root on Hong Yang farm, Qingpu district, be sub-packed in plastics bag after cleaning, preservation in-20 DEG C of refrigerators.Take out knee knurl during test, break into homogenate with plant tissue pulverizer, four layers of filtered through gauze, the centrifugal 10min of refrigerated centrifuge 4000r/min, abandons supernatant, and suspend with distilled water, repeat 3 times, being finally suspended into pathogen plasmodiophora concentration is 2 × 10
8individual/ml, obtained club root suspension, saves backup in 4 DEG C.
Be that the ratio mixing of 1:3 is mixed thoroughly with volume ratio by sterile soil and matrix, select the vinyl disc of water containing band concave-convex bottom, the middle foam cave dish placing 77 holes, sows Chinese cabbage seed in the dish of cave.Wherein, the main component of described matrix is vermiculite, peat and perlite, is purchased from Shanghai Sun Qiao modern agriculture Science and Technology Ltd..
Until Chinese cabbage seedling grow to three leaves wholeheartedly time, measure 200ml club root suspension with graduated cylinder, evenly water the seedling root being filled in each cave dish, club root suspension is flowed in vinyl disc by seedling, and club root suspension preserves 3d in vinyl disc.After 3d, Chinese cabbage seedling carries out cellar culture, daytime 28 DEG C, 10 hours, evening 18 DEG C, 14 hours, after cellar culture 45d, Chinese cabbage plant is dug out, clean root, whether the incidence of investigation plant, have knee knurl according to the root of Chinese cabbage and judge whether to have infected club root.See Fig. 2, the figure shows the Chinese cabbage infecting club root.See Fig. 3, the figure shows the Chinese cabbage not infecting club root.Statistics disease plant number, calculates sickness rate.Sickness rate=morbidity strain number/total strain number × 100%.See table 2, it is the qualification result of the anti-club root of different varieties Chinese cabbage.
The qualification result of the anti-club root of table 2 Chinese cabbage
As can be seen from Table 2: the Chinese cabbage of self-mating system material and intermediate materials is substantially all fallen ill, and the anti-club root Chinese cabbage cultivar of selected control material is not all fallen ill, illustrate that test meets anti-disease enzyme test requirements document substantially.In this test, 60 parts of self-mating system materials are except 13-1g-55 is not susceptible, and it is susceptible that other self-mating system materials all have in various degree, and what sickness rate was minimum is 13-1g-22 (21.4%); That sickness rate is the highest is 13-1g-8, up to 83%.The sickness rate of intermediate materials is also different, and wherein the sickness rate of 13-1g-69 and 13-1g-70 is lower, is respectively 27.3% and 21.7%; The sickness rate of 13-1g-65 is up to 61.5%.Illustrate that the anti-sense ability of each Chinese cabbage germ plasm resource to Cruciferae club root is different, fill with root authentication method by this kind and can obtain the induction reactance degree of each Chinese cabbage material to club root, for the anti-club root new variety of seed selection Chinese cabbage provide foundation.For intermediate materials, can according to different induction reactance abilities separation screening resistant material targetedly further, for the seed selection of Chinese cabbage anti-club root new variety provides more germ plasm resource.
Above are only part preferred embodiment of the present invention, the present invention is not limited in the content of embodiment.To those skilled in the art, can have various change and change in the concept of technical solution of the present invention, any change done and change, all within scope.
Claims (8)
1. an authentication method for the anti-club root of Chinese cabbage, the method comprises the steps:
Step 1, preparation pathogen plasmodiophora suspension;
Step 2, grows to three leaves wholeheartedly periods the seedling of Chinese cabbage, the club root suspension of aforementioned preparation is carried out uniform irrigation to seedling root;
Step 3, carries out cellar culture by the seedling after aforementioned impregnation, collects the Chinese cabbage not infecting club root.
2. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, is characterized in that: in described step 1, the concentration of the pathogen plasmodiophora suspension of preparation is 1 × 10
7~ 2 × 10
8individual/mL.
3. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, it is characterized in that, in described step 1, the compound method of club root suspension is: the fresh daetylorhiz collecting the Chinese cabbage of morbidity, in plant tissue crusher for crushing, filtration, centrifugal after cleaning, be modulated into pathogen plasmodiophora suspension with distilled water.
4. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, is characterized in that: after in described step 2, club root suspension is irrigated seedling root, and seedling root is flooded 3 ~ 4 days in club root suspension.
5. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, it is characterized in that, the type of rearing of described Chinese cabbage is: the vinyl disc selecting water containing band concave-convex bottom, in vinyl disc, place the foam cave dish with multiple hole, described Chinese cabbage sowing is grown in the dish of foam cave.
6. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 5, it is characterized in that, the mode of described Chinese cabbage inoculation club root is: when the Chinese cabbage in the dish of cave grow to three leaves wholeheartedly time, the seedling root pouring 200ml club root suspension coiled in each cave.
7. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, is characterized in that, in described step 3, the condition of seedling cellar culture is: daytime cultivates 10 hours under 28 DEG C of conditions, and cultivate 14 hours under 18 DEG C of conditions in the evening.
8. the authentication method of the anti-club root of a kind of Chinese cabbage as claimed in claim 1, is characterized in that: in described step 3, the time of seedling cellar culture is 35 ~ 45 days.
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| CN201410668162.2A CN104404124A (en) | 2014-11-20 | 2014-11-20 | Identification method for clubroot resistance of non-heading cabbages |
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Cited By (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543391A (en) * | 2016-02-05 | 2016-05-04 | 北京市农林科学院 | InDel molecular marker for identifying clubroot-resistant QTL (quantitative trait locus) located on Chinese cabbage A03 chromosome and application thereof |
| CN105850717A (en) * | 2016-04-20 | 2016-08-17 | 上海市农业科学院 | Method for acquiring non-heading Chinese cabbage clubroot resistance germplasm resources |
| CN105875151A (en) * | 2016-04-20 | 2016-08-24 | 上海市农业科学院 | Rapid identification method of Brassic campestris ssp. chinensis Makino waterlogging tolerance |
| CN106035059A (en) * | 2016-05-26 | 2016-10-26 | 西北农林科技大学 | Method for artificially synthesizing novel anti-clubroot brassica napus material |
| CN106119120A (en) * | 2016-06-20 | 2016-11-16 | 安徽省农业科学院作物研究所 | A kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof |
| CN106520908A (en) * | 2016-09-23 | 2017-03-22 | 湖北省农业科学院经济作物研究所 | Identification method for clubroot resistance of alpine radish at seedling stage |
| CN110178579A (en) * | 2019-07-02 | 2019-08-30 | 沈阳农业大学 | A kind of Cruciferae clubroot identification method |
| CN112301090A (en) * | 2020-11-03 | 2021-02-02 | 上海市农业科学院 | Identification method of downy mildew-resistant germplasm material of non-heading Chinese cabbage |
| CN113016510A (en) * | 2021-03-12 | 2021-06-25 | 上海应用技术大学 | Identification method for preventing clubroot of cauliflower vegetables |
| CN115581196A (en) * | 2022-10-28 | 2023-01-10 | 云南省农业科学院经济作物研究所 | Inoculation method for rapidly screening clubroot disease resistant varieties in seedling stage |
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Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105543391A (en) * | 2016-02-05 | 2016-05-04 | 北京市农林科学院 | InDel molecular marker for identifying clubroot-resistant QTL (quantitative trait locus) located on Chinese cabbage A03 chromosome and application thereof |
| CN105850717A (en) * | 2016-04-20 | 2016-08-17 | 上海市农业科学院 | Method for acquiring non-heading Chinese cabbage clubroot resistance germplasm resources |
| CN105875151A (en) * | 2016-04-20 | 2016-08-24 | 上海市农业科学院 | Rapid identification method of Brassic campestris ssp. chinensis Makino waterlogging tolerance |
| CN106035059A (en) * | 2016-05-26 | 2016-10-26 | 西北农林科技大学 | Method for artificially synthesizing novel anti-clubroot brassica napus material |
| CN106119120A (en) * | 2016-06-20 | 2016-11-16 | 安徽省农业科学院作物研究所 | A kind of plasmodiophora brassicae inoculation nutrient matrix and preparation method thereof |
| CN106520908A (en) * | 2016-09-23 | 2017-03-22 | 湖北省农业科学院经济作物研究所 | Identification method for clubroot resistance of alpine radish at seedling stage |
| CN110178579A (en) * | 2019-07-02 | 2019-08-30 | 沈阳农业大学 | A kind of Cruciferae clubroot identification method |
| CN110178579B (en) * | 2019-07-02 | 2021-09-24 | 沈阳农业大学 | Cruciferous clubroot identification method |
| CN112301090A (en) * | 2020-11-03 | 2021-02-02 | 上海市农业科学院 | Identification method of downy mildew-resistant germplasm material of non-heading Chinese cabbage |
| CN113016510A (en) * | 2021-03-12 | 2021-06-25 | 上海应用技术大学 | Identification method for preventing clubroot of cauliflower vegetables |
| CN115581196A (en) * | 2022-10-28 | 2023-01-10 | 云南省农业科学院经济作物研究所 | Inoculation method for rapidly screening clubroot disease resistant varieties in seedling stage |
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Application publication date: 20150311 |