CN106508336A - Identification method for clubroot disease resistance of high mountain radishes in field - Google Patents
Identification method for clubroot disease resistance of high mountain radishes in field Download PDFInfo
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- CN106508336A CN106508336A CN201610844634.4A CN201610844634A CN106508336A CN 106508336 A CN106508336 A CN 106508336A CN 201610844634 A CN201610844634 A CN 201610844634A CN 106508336 A CN106508336 A CN 106508336A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G7/00—Botany in general
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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Abstract
The invention discloses an identification method for clubroot disease resistance of high mountain radishes in a field. The identification method comprises the following steps: selecting experimental radish varieties, enriching bacterium sources of a clubroot disease nursery in a high mountain region, and sowing radishes; grading clubroot disease morbidity degree of full-grown radishes to obtain disease condition grades of the radishes; calculating radish variety disease condition indexes according to the disease condition grades of the radishes and a radish variety disease condition index general formula; and evaluating the resistance type of each radish variety to the clubroot disease by combining a radish disease resistance evaluation standard according to a calculation result. According to the identification method, the disease condition grading of the radishes which are infected with a disease is simplified; the influence on radish clubroot disease condition grading caused by black spots and black scars on the radishes is excluded; the existing grading mode is changed essentially; the overall identification process and the result processing analysis become simple, and meanwhile, the clubroot disease resistance of different radish varieties is judged rigorously and accurately.
Description
Technical field
The invention belongs to the anti-clubroot authentication method of crops, and in particular to a kind of mirror of the anti-clubroot in high chervil field
Determine method.
Background technology
Radix Raphani is 1～2 year raw herbaceous dicotyledon of Cruciferae Rhaphanuss, is the second largest vegetable of China, in China
There is the cultivation history of more than 2700 years, variety source is nearly 2000 parts, its variety source type very abundant, each ecological zone has suitable
Close traditional local varieties of one's respective area consumption habit and Various Seasonal growth.China Radix Raphani year sown area has exceeded 18,300,000 mu, entirely
There are plantation in each province of state, city, counties and districts, and the areas such as Japan, Hong Kong, Taiwan and Southeast Asia are simultaneously found a good sale in the Radix Raphani product year-round supply whole nation.
Clubroot is a kind of worldwide soil-borne disease, is found in Mediterranean two sides and south of europe earliest, and current many countries are all
There are generation, particularly Temperate Region in China.In recent years, warmed with global climate year by year, soil acidification degree increases, crop in cruciferae
Cultivated area constantly expands, the planting base vegetable production crop rotation time limit increases and the mutual allocation and transportation of north and south dish, crop in cruciferae root
Swollen disease occurring area expands rapidly, and the extent of injury also increases year by year, and clubroot has become one of important disease of crop in cruciferae.
Forefathers are found that Crr1, Crr2, Crr3, Crr4 by the genetic analyses to the anti-clubroot of Brassica genus Chinese cabbage,
CRa,CRb,CRk(SUWABE K,2003；SUWABE,2006；HIRAI M,2004；MATSUMOTO E,1998；SAKAMOTO
K,2008；PIAO Z Y,2002；PIAO Z Y, 2004) 7 anti-clubroot (clubroot resistance, CR) genes；SAITO
Deng (2006) by the qtl analysis to the anti-clubroot of arabidopsiss, precise positioning Crr3 genes；HATAKEYAMA etc. (2013) is big
Precise positioning Crr1 genes in Chinese cabbage；Piao etc. (2002,2004) has found CRb gene pairss clubroot biological strains by Chinese cabbage
2,4,8 show as resistance；CHIAUG etc. (1977) has found that anti-biological strain 1 and anti-biological strain 3 are by list in colea
One dominant gene is controlled, and anti-biological strain 5 is controlled by 2 pairs of recessive genes；CRUTE etc. (1983) have studied Caulis et Folium Brassicae campestriss pathogen plasmodiophora
Some bacterial strains and 3 kinds of brassica plant (Radix Brassicae rapae, Brassica campestris L, Caulis et Folium Brassicae capitatae) genotype relation after, it is believed that Brassica campestris L and Radix Brassicae rapae are to different bacterium
The resistance of strain is different, and they are probably to be controlled by oligogene.Piao (2007) etc. utilizes SSR marker TCR01 and TCR09,
By anti-clubroot CRb gene mapping, in Chinese cabbage A3 linkage groups and between labelling, the genetic distance is 2.9cM, is entered using the two labellings
Resistance transformation between row Chinese cabbage and Chinese cabbage, obtains the NIL of 9 ' BJN3 '.(2014) such as Zhang Ten g
According to Research foundation before, finely positioning is carried out, developed 2 Chinese cabbage SSR markers, respectively TCR79 and TCR108,
The genetic distance between labelling is 0.14cM, and these researchs provide theoretical base for the anti-clubroot molecular mark of Chinese cabbage
Plinth.Jonghoon Lee etc. (2015) utilize 78 Caulis et Folium Brassicae capitatae F of high-flux sequence GBS technique constructions2The high density collection of illustrative plates of colony,
QTLs positioning results show that the resistance locus to No. 4 biological strains are different from the site of the similar research of Chinese cabbage before this, and research is opened
Two different QTL regions are sent out related to the resistance of clubroot.BRADSHAW etc. (1997) is cultivated using Tetraploid method
The Radix Brassicae rapae resistant to clubroot, Caulis et Folium Brassicae capitatae are gone out；MANZANARES-DAULEUX etc. (2000) is by the disease-resistant base of European feeding Radix Raphani
Because being transferred on Chinese cabbage, the Chinese cabbage of out anti-clubroot is cultivated.Mountain region king's Chinese cabbage cultivar of Korea is high in Hubei at present
Mountain fire burns the anti-clubroot in level ground and shows excellent, and the anti-clubroot Caulis et Folium Brassicae capitatae for first just reaching shows excellent in Lichuan County, Hubei alpine region.
Chen Huihui etc. (2012) 2 Chinese cabbage SSR markers according to the research and development of Piao etc., respectively TCR13 and
TCR74, between labelling, the genetic distance is 0.18cM.Chen Longzheng (2015) etc. and is not tied using the Chinese cabbage germplasm for carrying CRb genes
Ball Chinese cabbage carries out inter-subspecies hybrid and backcrossing, designs special primer by existing SSR marker, obtains more simple and rapid
SCAR mark CRb-R-25 is applied to molecular marker assisted selection, creates with Chinese cabbage inter-subspecies hybrid in combination with Chinese cabbage
The anti-clubroot new germ plasm of Chinese cabbage processed.Guo Haifeng (2005) is also had found between disease-resistant Rape Genotypes and Radix Brassicae rapae genotype
There may be common gene.Zhejiang University Jin Jing etc. (2014) constructs SSR molecular marker genetic map and navigates to 1
Recessive clubroot Resistance QTL CRQTL1 of the LOD value more than 2.Yang Zheng etc. (2015) is to the anti-clubroot gene locis of Chinese cabbage
The molecular marker of CRa and CRb is identified that Preliminary Study specify that anti-clubroot gene C Ra contained by 78 parts of Chinese cabbage materials
And CRb.Yao Qiuju etc. (2015) resists resistance of the Chinese cabbage cultivar of clubroot in Henan Xinye and is identified, as a result table
Bright, different cultivars is had differences to the plasmodiophora brassicae resistance in Xinye area, wherein Y670, Y714, Y752 performance immunity.High hope etc.
(2015) 20 kinds of Brassica Napus clubroot resources are screened, are as a result shown, China is miscellaneous No. 9, China it is double No. 3 show as compared with
Strong anti-clubroot ability, can be used as the anti-clubroot breeding material of Brassica campestris L.The country has carried out anti-clubroot DABAI vegetable at present
The selection-breeding planted, in Hubei, governor's YANG-fire burns level ground and Lichuan alpine region, and the anti-clubroot Chinese cabbage of constituent parts shows as Beijing vegetable
The capital spring CR2 and Jing Chun CR3 performance at center is excellent, and 12CR159 performances in Qingdao academy of agricultural sciences are excellent, and Tianjin Ke Run vegetables institute day is white
CR15 performances are excellent.Our studies in China lack anti-source to head cabbage varieties at present, temporarily not anti-clubroot kind.Radix Raphani is to knee
The resistance of disease preferably, still suffers from generation relative to other crops, and interracial widely different, correlational study is carried out.
A big class of the Radix Raphani as Cruciferae, is endangered by clubroot, particularly alpine region, clubroot in 2003 deeply
In Hubei, governor Yang Xian burns level ground and breaks out and spread, and reaches 200hm2 to onset area in 2005, and have no harvest area 3.3hm2 (Wang Wei
It is red etc., 2013).Radish root tumefacting disease is Hubei governor sun area and Lichuan City homogeneous kernel arteries and veins Vegetable industrialization and peasant at present
The greatest problem for facing is increased income, is to solve Production requirement most efficient method from disease-resistant variety.
The content of the invention
In order to provide the radish varieties of anti-clubroot to alpine region, the invention provides a kind of anti-knee in high chervil field
The authentication method of disease, methods described are reliable and stable, and analysis mode is simply clear and definite, solves what alpine region radish root tumefacting disease brought
Yield and quality harm, optimize the selection-breeding of the anti-clubroot kind of alpine region Radix Raphani.
The technical scheme is that：A kind of authentication method of the anti-clubroot in high chervil field, it is characterised in that identification
Step is as follows：
S1, select for radish root tumefacting disease identification several radish varieties；
S2, alpine region select garden as clubroot sick nursery, spring by the end of April to the first tenday period of a month in May the clubroot disease
Susceptible Chinese cabbage is planted in garden, enriches the bacterium source of garden, continuously sows the susceptible Chinese cabbage several years；
If S3, by step S2 enrich good bacterium source clubroot sick nursery be divided into individual clubroot sick nursery area, every kind of radish varieties
It is seeded in each clubroot sick nursery area respectively, the concrete sowing time is then by the end of June to by the end of July；
S4, resistance of each radish varieties to clubroot in each clubroot sick nursery area was investigated in after planting the 55th～60 day.
Type of seeding in the S3 is：
1) sprinkling herbicide and preventing and treating subterranean pest-insect medicament before sowing, lower base fertilizer, bag ditch 1m ridgings, the high 20cm in ridge～
25cm, the long 7.5m in railway carriage or compartment face, single file or double row sowing, pitch-row are 40cm × 20cm；
2) 2～3 are sowed per hole when sowing, direct final singling during 5～6 leaf phase is not transplanted seedlings；
3) after planting field carries out normal management, imposes foliage fertilizer 2～3 times, and preventing and treating diamondback moth, Pieris rapae and downy mildew are soft
Maize ear rot, prevents and treats flea beetle after Radish Emerging.
The concrete mode for investigating the resistance of each radish varieties clubroot in each clubroot sick nursery area in the S4 is as follows：
1) in each clubroot sick nursery area, each radish varieties chooses some strains respectively, cleans, and observes the form of Radix Raphani；
2) the severity Scaling series of each Radix Raphani is judged according to Radix Raphani severity Scaling series criterion；
3) the respective disease index of each radish varieties in each clubroot sick nursery area is calculated according to disease index computing formula；
4) disease index of identical radish varieties in each clubroot sick nursery area is sought into arithmetic average, is commented according to disease resistance
The disease resistance of valency standard determination each radish varieties to clubroot.
The Radix Raphani severity Scaling series criterion is：
0 grade：Without root nodule on root system；
1 grade：There are 1～2 scattered root nodule, the root nodule diameter≤3mm on main root or fibrous root；
2 grades：There are 3 or more than 3 scattered root nodules, 3mm ＜ root nodule diameters≤5mm on main root or fibrous root；
3 grades：On main root or on fibrous root, several root nodules are assembled, the root nodule diameter ＞ 5mm；
The disease index computing formula is：
The diseased plant number at different levels is severity Scaling series identical strain number in same radish varieties in each clubroot sick nursery area；
The investigation total strain number is the total strain number of same radish varieties in each clubroot sick nursery area；
The disease index is the extent of injury of the radish varieties by the clubroot；
The disease resistance evaluation standard is as follows：
Disease index is disease-resistant (R) 0～11.11；Disease index is resistance to disease (T) 11.12～33.33；Disease index
It is susceptible (S) 33.34～55.55；Disease index is high sense (HS) 55.56～100.00.
The clubroot sick nursery is covered with mulch film when planting each radish varieties.
The bacterium source is all from the plasmodiophora brassicae of the alpine region.
Beneficial effects of the present invention are：The authentication method of the relatively existing radish root tumefacting disease of authentication method of the present invention
It is simpler, judge only up to 3 grades of the severity Scaling series of radish root tumefacting disease symptom, the criterion of severity Scaling series is more
Apparent, the root nodule state completely on Radix Raphani is defined, and not only greatly reduces the workload and subsequent result analysis in qualification process
Process, while the accurate disease index of each radish varieties, can accurately judge the anti-clubroot ability of radish varieties.The present invention exists
On the basis of simplifying existing clubroot authentication method, reach than the more preferable effect of existing authentication method.
Description of the drawings
Fig. 1 is Radix Raphani severity Scaling canonical reference figure.
In Fig. 1, a-0 levels；B-1 levels；C-2 levels；D-3 levels.
Specific embodiment
Below in conjunction with specific embodiment, the present invention will be further described.
The clubroot harm of Yichang City, Hubei Province Chang Yang counties is serious, and it is each kind radish root tumefacting disease to select Chang Yang counties and burn level ground
The proving ground of identification, clubroot bacterium source are all from the plasmodiophora brassicae in Chang Yang counties, and annual spring is by the end of April to the first tenday period of a month in May in test base
The susceptible Chinese cabbage of ground plantation, at the beginning of 2016, optional test base has been the abundant clubroot sick nursery of bacterium source；With lake
Used as the radish varieties for preparing identification, test is shared the radish varieties breeding resources that Bei Sheng academies of agricultural sciences industrial crops institute is provided
155 radish varieties, are denoted as kind 1 to kind 155.
Above-mentioned clubroot sick nursery is divided into into Liang Ge areas, respectively A areas and B areas, in by the end of June, 2016 to by the end of July, in each area
Above-mentioned 155 radish varieties are sowed respectively, sowing step is as follows：
1) sprinkling herbicide and preventing and treating subterranean pest-insect medicament before sowing, lower base fertilizer, bag ditch 1m ridgings, the high 20cm in ridge～
25cm, the long 7.5m in railway carriage or compartment face, are covered with mulch film, single file or double row sowing, and pitch-row is 40cm × 20cm；
2) 2～3 are sowed per hole when sowing, 5～6 leaf phases direct final singling after emerging is not transplanted seedlings；
3) after planting field carries out normal management, imposes foliage fertilizer 2～3 times, and preventing and treating diamondback moth, Pieris rapae and downy mildew are soft
Maize ear rot, prevents and treats flea beetle after Radish Emerging；
Investigated within after planting the 55th～60 day, in investigation clubroot sick nursery A area and B areas, each radish varieties clubroot is anti-
Property, concrete investigation method is as follows：
1) all individual plants of each radish varieties in clubroot sick nursery A area are extracted, removes mud, observe the form of Radix Raphani；By root
The all individual plants of each radish varieties in swollen disease sick nursery B area are extracted, and are removed mud, are observed the form of Radix Raphani；
2) the morbidity series of each Radix Raphani is judged according to Radix Raphani severity Scaling standard, and logon data is recorded in retention of taking pictures；
3) disease index of 155 radish varieties in clubroot sick nursery A area is calculated according to disease index computing formula；Root
The disease index of 155 radish varieties in clubroot sick nursery B area is calculated according to disease index computing formula；
The Radix Raphani severity Scaling series criterion：
0 grade as shown in a figures in Fig. 1：Without root nodule on root system；
1 grade as shown in b figures in Fig. 1：There are 1～2 scattered root nodule, the root nodule diameter≤3mm on main root or fibrous root；
2 grades as shown in c figures in Fig. 1：There are 3 or more than 3 scattered root nodules, 3mm ＜ root nodule diameters≤5mm on main root or fibrous root；
3 grades as shown in d figures in Fig. 1：On main root or on fibrous root, several root nodules are assembled, the root nodule diameter ＞ 5mm；
Disease index computing formula：
During the diseased plant number at different levels is every piece of clubroot sick nursery, same radish varieties severity Scaling series identical strain number；
During the investigation total strain number is every piece of clubroot sick nursery, the total strain number of same radish varieties；
The disease index is the extent of injury of the radish varieties by the clubroot.
4) clubroot sick nursery A area is sought into arithmetic average with the disease index of identical radish varieties in B areas, according to disease resistance
Evaluation criterion judges disease resistance of 155 radish varieties to clubroot；
The disease resistance evaluation standard is as follows：
1 disease resistance evaluation standard of table
| Disease index | Evaluation of resistance |
| 0～11.11 | Disease-resistant (R) |
| 11.12～33.33 | Resistance to disease (T) |
| 33.34～55.55 | Susceptible (S) |
| 55.56～100 | High sense (HS) |
The qualification result of the anti-clubroot of above-mentioned 155 radish varieties is as follows：
The qualification result of the anti-clubroot of 2 radish varieties of table
From Table 2, it can be seen that resistance of level ground field clubroot 155 radish varieties of Disease garden identification to clubroot is burnt, disease
Feelings index is disease-resistant variety 0～11.11, and disease-resistant variety has 24 parts, and disease index is disease tolerant variety 11.12～33.33, resistance to
Sick kind has 34 parts.
From above-described embodiment as can be seen that the authentication method of the anti-clubroot in high chervil field of the present invention is simply easy
OK, on the disease index for judging to catch an illness Radix Raphani, eliminating the Radix Raphani black speck in existing clubroot authentication method and black scar affects,
Although the black speck on Radix Raphani and scar occur after Radix Raphani is infected by clubroot, there is presently no strong evidence can
It can be that plasmodiophora brassicae is seen in grinding under the microscope that the pathogen infected with proof is exactly necessarily the root nodule on plasmodiophora brassicae, but Radix Raphani
Spore, and spore can be with renewed vaccination on new plant, with this, forming same root nodule proves that plasmodiophora brassicae causes Radix Raphani
Root nodule is generated, only using the form and quantity of root nodule as the criterion of Radix Raphani severity Scaling of catching an illness, whole identification is enormously simplify
Process, or even the result for judging is more rigorous accurately, inherently for providing new method in the identification of radish root tumefacting disease.
Claims (8)
1. the authentication method of the anti-clubroot in a kind of high chervil field, it is characterised in that authentication step is as follows：
S1, select for radish root tumefacting disease identification several radish varieties；
S2, alpine region select garden as clubroot sick nursery, spring is by the end of April to the first tenday period of a month in May in the clubroot sick nursery kind
Susceptible Chinese cabbage is planted, the bacterium source of garden is enriched, is continuously sowed the susceptible Chinese cabbage several years；
S3, the clubroot sick nursery for enriching good bacterium source in step S2 is divided into into several clubroot sick nursery areas, every kind of radish varieties point
It is not seeded in each clubroot sick nursery area, the concrete sowing time is then by the end of June to by the end of July；
S4, the resistance for investigating each radish varieties clubroot in each clubroot sick nursery area in after planting the 55th to 60 day.
2. a kind of authentication method of the anti-clubroot in high chervil field according to claim 1, it is characterised in that the S3
Middle type of seeding is：
1) sprinkling herbicide and preventing and treating subterranean pest-insect medicament, lower base fertilizer, bag ditch 1m ridgings, the high 20cm～25cm in ridge, railway carriage or compartment before sowing
The long 7.5m in face, double row sowing, pitch-row are 40cm × 20cm；
2) 2～3 are sowed per hole when sowing, 5～6 leaf phases, direct final singling was not transplanted seedlings；
3) after planting field carries out normal management, imposes foliage fertilizer 2～3 times, preventing and treating diamondback moth, Pieris rapae and the soft corruption of downy mildew
Disease, prevents and treats flea beetle after Radish Emerging.
3. a kind of authentication method of the anti-clubroot in high chervil field according to claim 1, it is characterised in that the S4
The middle concrete mode for investigating the resistance of each radish varieties clubroot in each clubroot sick nursery area is as follows：
1) in each clubroot sick nursery area, each radish varieties chooses some strains respectively, cleans, and observes the form of Radix Raphani；
2) the sick level of each Radix Raphani is judged according to Radix Raphani severity Scaling series criterion；
3) the respective disease index of each radish varieties in each clubroot sick nursery area is calculated according to disease index computing formula；
4) disease index of identical radish varieties in each clubroot sick nursery area is sought into arithmetic average, according to disease resistance evaluation mark
It is accurate to judge disease resistance of each radish varieties to clubroot.
4. a kind of authentication method of the anti-clubroot in high chervil field according to claim 3, it is characterised in that the Radix Raphani
Severity Scaling series criterion is：
0 grade：Without root nodule on root system；
1 grade：There are 1～2 scattered root nodule, the root nodule diameter≤3mm on main root or fibrous root；
2 grades：There are 3 or more than 3 scattered root nodules, 3mm ＜ root nodule diameters≤5mm on main root or fibrous root；
3 grades：On main root or on fibrous root, several root nodules are assembled, the root nodule diameter ＞ 5mm.
5. a kind of authentication method of the anti-clubroot in high chervil field according to claim 3, it is characterised in that the disease
Feelings formula of index is：
The diseased plant number at different levels is severity Scaling series identical strain number in same radish varieties in each clubroot sick nursery area；
The investigation total strain number is the total strain number of same radish varieties in each clubroot sick nursery area；
The disease index is the extent of injury of the radish varieties by the clubroot.
6. the authentication method of the anti-clubroot in a kind of high chervil field according to claim 3, it is characterised in that described anti-
Characteristic of disease evaluation criterion is as follows：
Disease index is disease-resistant (R) 0～11.11；Disease index is resistance to disease (T) 11.12～33.33；Disease index exists
33.34～55.55 is susceptible (S)；Disease index is high sense (HS) 55.56～100.00.
7. the authentication method of the anti-clubroot in a kind of high chervil field according to claim 1, it is characterised in that：Described
Swollen disease sick nursery is covered with mulch film when planting each radish varieties.
8. a kind of authentication method of the anti-clubroot in high chervil field according to claim 1, it is characterised in that the bacterium
Source is all from the plasmodiophora brassicae of the alpine region.
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| CN108575645A (en) * | 2018-04-03 | 2018-09-28 | 中国农业科学院棉花研究所 | A kind of method of field Fast Evaluation cotton to herbicide tolerant |
| CN108739147A (en) * | 2018-06-13 | 2018-11-06 | 山东省潍坊市农业科学院 | A method of the anti-clubroot Chinese cabbage germplasm of initiative |
| CN108739147B (en) * | 2018-06-13 | 2020-09-01 | 山东省潍坊市农业科学院 | Method for creating clubroot-resistant Chinese cabbage germplasm |
| CN109526443A (en) * | 2019-01-14 | 2019-03-29 | 长江师范学院 | A kind of rapid identification method of radish disease resistance |
| CN109526443B (en) * | 2019-01-14 | 2020-12-29 | 长江师范学院 | Method for rapidly identifying disease resistance of radish |
| CN109699357A (en) * | 2019-01-31 | 2019-05-03 | 南京信息工程大学 | A kind of facility grape downy mildew application method |
| 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 |
| CN113234846A (en) * | 2021-05-09 | 2021-08-10 | 湖北省农业科学院经济作物研究所 | Radish clubroot-resistant molecular marker and application thereof |
| CN113234846B (en) * | 2021-05-09 | 2022-02-01 | 湖北省农业科学院经济作物研究所 | Radish clubroot-resistant molecular marker and application thereof |
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