CN100446655C - Seedling stage identification method for tobacco black shank resistance - Google Patents
Seedling stage identification method for tobacco black shank resistance Download PDFInfo
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Abstract
The invention discloses a tobacco black shank resistance seedling stage identification method, which separates pathogenic bacteria from susceptible tobacco stems, and obtains a black shank inoculum after purification and culture. And (3) inoculating the inoculum to the stem base part of the tobacco seedling to be tested, recording disease fingers once respectively on the 5 th day, the 7 th day and the 10 th day after inoculation, and taking the average value of the disease fingers three times as the resistance index of the tested variety. The method provided by the invention firstly realizes the replacement of the traditional field identification result with the seedling stage identification result, effectively avoids the influence of the external environment on the identification result, and improves the accuracy of the tobacco black shank resistance identification. The method only needs about 65 days from seedling culture to identification completion, and greatly shortens the identification time. The method is simple and convenient to operate, low in cost, real and reliable in identification result and good in market application prospect.
Description
Technical field
The invention belongs to disease resistance of plant authenticate technology field, be specifically related to the evaluation of black shank resistance, particularly relate to a kind of method of the black shank resistance being carried out the evaluation in seedling stage.
Background technology
Black shank is commonly called as " waist mashed ", " wearing unlined long gown ", " sending out pest ", " underground disease " etc., is a kind of widely distributed, tobacco diseases that harm is serious.Be found in the Java of Indonesia first in 1896, after nineteen twenty-four, all there is this disease report in the product cigarette district in temperate zone, the whole world, subtropics and the torrid zone, has only Zimbabwe still not have record so far.Nineteen fifty, China found black shank first in the Yellow River and Huai He River cigarette district.At present, China's morbidity crop area every year on average is up to 76373hm
2, direct economic loss is above 1.23 hundred million yuans.Therefore, how to improve the resistance against diseases of existing tobacco bred, cultivate tobacco bred, just become the focus of this area research at present with high resistance against diseases.
As the key of breed breeding, existing tobacco resistance authentication method mainly contains indoor excised leaf inoculation method and spontaneous induction resistance survey method.Though indoor excised leaf inoculation method is simple, the result of its evaluation can only reflect the anti-extended capability of excised leaf to disease, can't reflect the actual resistance of kind, can only be as the reference index of resistance division.Though spontaneous induction resistance authentication method can reflect the natural resistance against diseases of kind, this method is subject to planting conditions and climatic influences, and the result that same kind is identified in different cigarettes district can exist than big-difference.Simultaneously, because black shank is very responsive to temperature and humidity conditions, only (28～32 ℃, relative moisture more than 80%) could rapid spread under high temperature, high humidity environment.When the environmental condition between the test time was unfavorable for that disease takes place, this method can't obtain measurement result exactly.
The manual-induced authentication method in disease garden, land for growing field crops that occurs subsequently, though by setting up the disease garden, artificial quantitatively inoculation pathogen, means such as simulating nature onset condition have improved the accuracy of test, but still can't fundamentally solve the influence of external environment condition to qualification result.Simultaneously, factors such as the amount of pathogen inoculation and inoculation time all can exert an influence to final result.Therefore, this method also exists identification of species and limited amount, and problems such as fluctuation easily take place qualification result.And finishing of identifying from growing seedlings to of this method need be nearly 195 days, and qualification time is long.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of sprout period testifying method of black shank resistance is provided, with the alternative traditional land for growing field crops of qualification result in seedling stage qualification result, avoid the influence of external environment condition to qualification result, improve the accuracy that the black shank resistance is identified, shorten qualification time.
Purpose of the present invention is achieved by the following technical programs.
A kind of sprout period testifying method of black shank resistance comprises the step of following order:
1. pathogen separates
Adding content in basal medium is the rifampin solution of 3000ug/ml, and it is standby to make the rifampin medium; Gather the typical ill tobacco stem of black shank symptom, get disease stem epidermis hypochondriasis and organize several piece, be cut into the fritter of long 1～2cm, place on the rifampin medium, behind cultivation, purifying, obtain the black shank mycelium, at ambient temperature, be stored in the sterile purified water standby;
Described basal medium is an oat medium.
The volume percent content of rifampin stoste is 1.3% in the described rifampin medium.
2. the preparation of balck shank inoculum
With the black shank mycelium move into cultivate 5～7 days in the oat culture fluid after, change over to cultivate on the paddy medium and obtained the balck shank inoculum in 18～20 days;
The preparation method of described paddy medium is: the millet immersion was cleaned with clear water after 6 hours, wash in a pan shrivelled ghost grain to the greatest extent, boil and blow into half-open flower-shapedly to grain, it is that 50%～60% back is divided in the triangular flask of packing into that cooling is dried in the air to the millet water content, and 115 ℃ of following autoclavings 1 hour promptly;
The grain amount of packing in the triangular flask is advisable to be no more than 3/4 bottle;
3. design of sick garden and management
To the greenhouse disease garden, the cigarette seedling is randomized arrangement in sick garden with 45～55 days cigarette transplantation of seedlings to be identified of seedling age, establishes 3 repetitions, whenever repeats to plant cigarette 10 strains, and greenhouse management technology routinely manages; With little gold 1025 is susceptible contrast, and the big gold dollar of safflower is susceptible auxiliary contrast, and Venus 6007 resists contrast in being, reforming No. 3 is disease-resistant contrast;
4. bacterium paddy inoculation
After the cigarette transplantation of seedlings 5～7 days, will cultivate the basal part of stem that the inoculum that obtains is seeded to cigarette seedling to be measured through step (2), inoculum concentration is the 0.5g/ strain; The earthing and the 15～20cm that pours water immediately after the inoculation kept 24～48 hours.
5. identify and classification
In the greenhouse disease garden, write down respectively once respectively at postvaccinal the 5th day, the 7th day and the 10th day that disease refers to, the mean value that refers to three diseases is as the resistance index of test kind; And then identification of species is carried out classification by national standard.
The present invention takes the lead in proposing a kind of black shank sprout period testifying method with practical value, realized with the alternative traditional land for growing field crops of qualification result in seedling stage qualification result, effectively avoid the influence of external environment condition, improved the accuracy that the black shank resistance is identified qualification result.Test shows that only finishing from the evaluation of growing seedlings needs about 65 days, has shortened qualification time significantly.This method is easy and simple to handle, and is with low cost, and qualification result is true and reliable, has better market prospect.
Description of drawings
Fig. 1 refers to changing trend diagram for 5,7,10 days disease of different resistant variety inoculation balck shanks.
Embodiment
The present invention is further detailed explanation below in conjunction with embodiment and accompanying drawing, but they are not limitation of the invention.
Embodiment 1
Get 21 of medical rifampicin capsules and add 95% ethanol 100ml, and add the extremely dissolving of 1NNaOH number, make the rifampin stoste of 3000ug/ml (rifampin content), place 4 ℃ of refrigerators to preserve.Add 2ml rifampin stoste and mix in every 150ml oat medium, it is standby to make the rifampin medium.Gather the typical ill tobacco stem of black shank symptom, get disease stem epidermis hypochondriasis and organize several piece, be cut into the fritter of long 1cm, place on the rifampin medium, behind cultivation, purifying, obtain the black shank mycelium, at ambient temperature, be stored in the sterile purified water standby.The millet immersion was cleaned with clear water after 6 hours, wash in a pan shrivelled ghost grain to the greatest extent, boil blow into to grain half-open flower-shaped, it is that 50% back is divided in the 500ml triangular flask of packing into that cooling is dried in the air to the millet water content, the grain amount of packing into is advisable to be no more than the 500ml graduation mark, and it is standby to make bacterium paddy medium in 1 hour at 115 ℃ of following autoclavings.With the black shank mycelium move into cultivate 5 days in the oat culture fluid after, change over to again to cultivate on the bacterium paddy medium and obtained the balck shank inoculum in 18 days.To the greenhouse disease garden, the cigarette seedling is randomized arrangement in sick garden with 50 days cigarette transplantation of seedlings to be identified of seedling age, establishes 3 repetitions, whenever repeats to plant cigarette 10 strains, and greenhouse management technology routinely manages.With little gold 1025 is susceptible contrast, and the big gold dollar of safflower is susceptible auxiliary contrast, and Venus 6007 resists contrast in being, reforming No. 3 is disease-resistant contrast.After the cigarette transplantation of seedlings 5 days, inoculum is seeded to the basal part of stem of cigarette seedling to be measured, inoculum concentration is the 0.5g/ strain.The earthing and the 15cm that pours water immediately after the inoculation kept 24 hours.In the greenhouse disease garden, write down respectively once respectively at postvaccinal the 5th day, the 7th day and the 10th day that disease refers to, the mean value that refers to three diseases is as the resistance index of test kind.By national sector standard YC/T39-1996 the identification of species resistance is divided into level Four: disease-resistant (R): disease refers to below 25; In anti-(MR); Disease refers at 25.1-50.0; Middle sense (MS): disease refers at 5 0.1-75.0; Susceptible (S): disease refers to more than 75.
Embodiment 2
Repeat embodiment 1, following difference is arranged: the black shank mycelium was cultivated 6 days in the oat culture fluid.
Embodiment 3
Repeat embodiment 1, following difference is arranged: the black shank mycelium was cultivated 7 days in the oat culture fluid.
Embodiment 4
Repeat embodiment 1, following difference is arranged: the black shank mycelium was cultivated 19 days in bacterium paddy medium.
Embodiment 5
Repeat embodiment 1, following difference is arranged: the black shank mycelium was cultivated 20 days in bacterium paddy medium.
Embodiment 6
Repeat embodiment 1, following difference arranged: with 45 days cigarette transplantation of seedlings to be identified of seedling age to the greenhouse disease garden.
Embodiment 7
Repeat embodiment 1, following difference arranged: with 55 days cigarette transplantation of seedlings to be identified of seedling age to the greenhouse disease garden.
Embodiment 8
Repeat embodiment 1, following difference is arranged: the cigarette transplantation of seedlings was seeded to inoculum the basal part of stem of cigarette seedling to be measured after 6 days.
Embodiment 9
Repeat embodiment 1, following difference is arranged: the cigarette transplantation of seedlings was seeded to inoculum the basal part of stem of cigarette seedling to be measured after 7 days.
Repeat embodiment 1, following difference arranged: well-done millet cooling dry in the air to water content be 55%.
Embodiment 11
Repeat embodiment 1, following difference arranged: well-done millet cooling dry in the air to water content be 60%.
Embodiment 12
Repeat embodiment 1, following difference is arranged: the earthing and the 18cm that pours water immediately after the inoculation kept 36 hours.
Embodiment 13
Repeat embodiment 1, following difference is arranged: the earthing and the 20cm that pours water immediately after the inoculation kept 48 hours.
Experimental example 1
1. experiment condition
1. tested variety: 27 parts of kinds of the examination of whole nation district, the examination of provinces and regions, Yunnan, see table 1 for details.With little gold 1025 is susceptible contrast, and the big gold dollar of safflower is susceptible auxiliary contrast, and Venus 6007 resists contrast in being, reforming No. 3 is disease-resistant contrast.
2. test the place: the sick garden of greenhouse, Agriculture institute, Yunnan tobacco sciences base balck shank.
3. the preparation of balck shank inoculum: the black shank bacterial strain (being numbered P0402) of separation and purification is moved in the oat culture fluid shaking table cultivates, change over to after 7 days cultivate on the bacterium paddy medium 18 days standby.The making of bacterium paddy: millet soaked clean with clear water after 6 hours, boil and blow into half-open flower-shapedly to grain, cooling is dried in the air to half-dried, divides 1 hour (115 ℃) of 500ml triangular flask mesohigh sterilization of packing into.The grain amount of packing into is advisable to be no more than triangular flask 500ml graduation mark.
4. experimental design of balck shank greenhouse disease garden and management: each identification of species is in greenhouse disease garden randomized arrangement, repeats for 3 times, whenever repeats to plant cigarette 10 strains, and technology is carried out greenhouse management routinely.
5. balck shank inoculation: 50 days cigarette transplantation of seedlings of seedling age to the greenhouse disease garden, is transplanted inoculation bacterium paddy after back 5 days, the bacterium paddy of cultivating 18 days is seeded to the basal part of stem of cigarette strain, inoculum concentration is the 0.5g/ strain.Earthing and preserving moisture immediately after the inoculation makes that soil is in saturated or hypersaturated state in the disease garden.
6. identify: respectively investigated one time the state of an illness in 5,7,10 days in the inoculation back, the mean value that refers to three diseases is as the resistance index of test kind.
7. classification: by national sector standard YC/T39-1996 the resistance of kind is divided into level Four: disease-resistant (R): disease refers to below 25; In anti-(MR); Disease refers at 25.1-50.0; Middle sense (MS): disease refers at 50.1-75.0; Susceptible (S): disease refers to more than 75.
8. the contrast experiment organizes: adopt artificial induction land for growing field crops strain-forming period resistance authentication method, bacterium paddy inoculum concentration is the 5g/ strain, and carries out wound and the inoculation test of non-wound respectively in field, two sick gardens, and qualification result and seedling stage are identified contrast, see table 1 for details.Stab the stem foot epidermis with aseptic pocket knife when wherein the wound test is meant inoculation.
Table 1 balck shank seedling stage and the contrast of land for growing field crops strain-forming period resistance qualification result
Continuous table 1:
2. interpretation of result: greenhouse qualification result and the land for growing field crops not correlation coefficient of the qualification result of wound inoculation are R
2=0.9609, the correlation coefficient between the qualification result of greenhouse qualification result and the inoculation of land for growing field crops wound is R
3=0.8148, look into the significance of correlation coefficient schedule, R
1, R
2, R
3On 1% level, all reach remarkable positive correlation, and the correlation highly significant.
From above qualification result as can be seen, when seedling stage, inoculum concentration was the 0.5g/ strain and the qualification result of land for growing field crops inoculum concentration when being the 5g/ strain reach unanimity, can replace the land for growing field crops qualification result with the qualification result in seedling stage fully.
Experimental example 2
1. experiment condition
1. tested variety: 50 parts of germ plasm resources that this project team provides, variety name sees Table 2.
2. test period and place: 2004～2005 years in the sick garden of greenhouse, Yunnan Provine Tobacco Science Inst. base balck shank.
3. disease resistance authentication method
Each kind is all established 3 repetitions, every repetition 10 strains, randomized arrangement.With Xiao Huang 1025 is susceptible contrast, and the big gold dollar of safflower is susceptible auxiliary contrast, and Venus 6007 resists contrast in being, reforming No. 3 is resistant control.Adopt basal part of stem bacterium paddy inoculation method during evaluation, with 50 days cigarette transplantation of seedlings of seedling age to the greenhouse disease garden, transplant inoculation in back 5 days, the bacterium paddy of cultivating 18 days is inoculated in the basal part of stem of cigarette seedling by the inoculum concentration of 0.5g/ strain, and in time earthing is preserved moisture, inoculation back is respectively at the 5th day, the 7th day and the 10th day investigation state of an illness, refers to as the resistance criteria for classifying with the average disease of investigation in 5,7,10 days.
4. disease and resistance grade scale
During disease survey, degree of being in a bad way classification is carried out by national sector standard YC/T39-1996 regulation; According to the incidence of disease severity and check variety, refer to that by the last disease of determining generally disease is divided into following level Four: disease-resistant (R): disease refers to below 25; In anti-(MR); Disease refers at 25.1-50.0; Middle sense (MS): disease refers at 50.1-75.0; Susceptible (S): disease refers to more than 75.
2. result and analysis
1. the anti-balck shank qualification result of germ plasm resource
Qualification result shows: anti-balck shank (R) germplasm has 13 parts, in anti-balck shank (MR) germplasm have 25 parts; In 50 parts of materials, have 76% germ plasm resource during the resistance level of balck shank is shown as anti-more than; Have 10 parts of kinds this disease of verifying to show as middle sense (MS), 2 parts of germplasms show as susceptible (S), and the ratio that susceptible variety accounts for is less.Qualification result sees table 2 and table 3 for details.
The anti-balck shank qualification result of table 2 germ plasm resource
| | 5d | 7d | 10d | Average disease refers to | Evaluation of resistance | ||
| 2017 | 17.50 | 28.33 | 31.67 | 25.83 | MR | ||
| 9908 | 29.17 | 36.67 | 43.33 | 36.39 | MR | ||
| 99K521 | 19.17 | 26.67 | 46.67 | 30.83 | MR | ||
| 99F112 | 26.67 | 29.17 | 37.50 | 31.11 | MR | ||
| Hubei Province cigarette 1# | 26.67 | 35.00 | 42.50 | 34.72 | MR | ||
| TN86 | 11.67 | 35.00 | 35.83 | 27.50 | MR | ||
| 2009 | 25.00 | 32.50 | 48.75 | 35.42 | MR | ||
| Fragrant No. 1 of cloud | 25.83 | 30.83 | 34.17 | 30.28 | MR | ||
| No. 11, Bath agate | 30.00 | 40.83 | 50.83 | 40.56 | MR | ||
| No. 12, Bath agate | 13.33 | 25.00 | 43.33 | 27.22 | MR | ||
| H2 | 21.02 | 40.28 | 55.00 | 38.77 | MR | ||
| H4 | 20.00 | 21.67 | 35.83 | 25.83 | MR | ||
| H5 | 17.50 | 43.33 | 52.50 | 37.78 | MR | ||
| H6 | 34.17 | 39.17 | 50.00 | 41.11 | MR | ||
| H7 | 30.83 | 53.33 | 64.17 | 49.44 | MR | ||
| H12 | 24.17 | 46.67 | 56.67 | 42.50 | MR | ||
| H16 | 32.50 | 36.67 | 50.00 | 39.72 | MR | ||
| H17 | 25.83 | 48.33 | 58.33 | 44.17 | MR | ||
| MC1 | 32.50 | 35.83 | 50.00 | 39.44 | MR | ||
| No. 5, platform cigarette | 38.33 | 46.67 | 56.67 | 47.22 | MR | ||
| Carifoniavalle | 23.33 | 26.67 | 34.17 | 28.06 | MR | ||
| Islangold | 33.33 | 40.83 | 45.45 | 39.87 | MR | ||
| 8022 | 24.17 | 33.33 | 43.33 | 33.61 | MR | ||
| The Xiao Bai muscle | 38.33 | 45.00 | 50.00 | 44.44 | MR | ||
| Crust draws No. 1 | 25.00 | 25.83 | 30.83 | 27.22 | MR | ||
| 50926 | 60.83 | 66.67 | 71.67 | 66.39 | MS |
Continuous table 2:
| YNBS1 | 49.17 | 67.50 | 69.17 | 61.94 | MS |
| H1 | 35.83 | 49.17 | 72.31 | 52.44 | MS |
| H8 | 45.83 | 75.00 | 77.50 | 66.11 | MS |
| H10 | 40.00 | 64.17 | 72.50 | 58.89 | MS |
| H11 | 45.83 | 55.00 | 58.33 | 53.06 | MS |
| H13 | 38.52 | 60.00 | 68.33 | 55.62 | MS |
| H15 | 43.33 | 60.00 | 75.00 | 59.44 | MS |
| Huang Miao | 45.83 | 56.67 | 67.50 | 56.67 | MS |
| Candel | 57.50 | 70.00 | 77.50 | 68.33 | MS |
| Pan Yuanhuang | 12.69 | 16.67 | 23.33 | 17.56 | R |
| H80A432 | 4.17 | 10.83 | 11.67 | 8.89 | R |
| Coker Tobacco | 6.67 | 6.67 | 7.50 | 6.94 | R |
| Land Huang | 2.50 | 5.00 | 8.33 | 5.28 | R |
| Peace is selected 79-1 | 5.00 | 5.83 | 10.83 | 7.22 | R |
| No. 7, platform cigarette | 20.00 | 24.17 | 25.83 | 23.33 | R |
| Build ripple No. 1 | 0.00 | 0.83 | 3.33 | 1.39 | R |
| NC100 | 19.17 | 20.83 | 23.33 | 21.11 | R |
| Draw No. 3 in Tianjin | 7.50 | 15.83 | 22.50 | 15.28 | R |
| OX414 | 5.00 | 6.67 | 12.50 | 8.06 | R |
| Draw No. 1 in Tianjin | 0.00 | 2.50 | 5.00 | 2.50 | R |
| The COM cigarette | 4.17 | 4.17 | 13.33 | 7.22 | R |
| Crust draws No. 2 | 10.00 | 13.33 | 15.83 | 13.06 | R |
| H9 | 70.83 | 72.50 | 89.72 | 80.28 | S |
| N.Syivestris | 75.00 | 82.50 | 85.00 | 80.83 | S |
| Little gold 1025 | 68.33 | 89.17 | 93.33 | 83.61 | S |
| Venus 6007 | 20.00 | 55.00 | 60.00 | 45.00 | MR |
| Reform No. 3 | 12.50 | 21.67 | 35.00 | 23.06 | R |
| The big gold dollar of safflower | 67.50 | 75.00 | 85.00 | 75.83 | S |
Table 3 is for the disease resistance qualification result statistical form of the germ plasm resource of trying to the black shank disease
| The resistance level resistance is identified | Disease-resistant variety R | In anti-kind M R | Middle sense kind M S | Susceptible variety S | Kind amounts to |
| Kind number (individual) | 13 | 25 | 10 | 2 | 50 |
| Different resistance level kind proportions (%) | 26％ | 50％ | 20％ | 4％ | 100％ |
2. the disease of different resistant variety inoculation balck shanks after 5 days, 7 days, 10 days refers to change.
The the 5th, 7,10 day sick fingering row comparative analysis is found: when referring to refer to subtract each other with the 5th day disease with the 7th day disease, both diseases refer to that difference accounts for 66% of tested variety in the kind below 10, wherein disease-resistant variety (R) has 13 parts, accounts for 100% of disease-resistant variety sum; In anti-kind (MR) have 16 parts, in accounting for 64% of anti-kind sum; Middle sense kind (MS) has 2 parts, feels 20% of kind sum in accounting for, and susceptible variety (S) has 2 parts, accounts for 100% of susceptible variety sum.Difference accounts for 34% of tested variety in the kind more than 10, and anti-in being, middle sense kind, wherein in anti-kind (MR) have 9 parts, in accounting for 36% of anti-kind sum; Middle sense kind (MS) has 8 parts, feels 80% of kind sum in accounting for.
Whole disease-resistant variety (R) and susceptible variety (S) refer to that the 5th～7 day disease difference is all below 10; Feel in the kind and difference is anti-and middle in more than 10, though in anti-and in the sense kind suitable, but there is 80% middle sense kind disease in this is identified to refer to that difference is more than 10, the sense kind is relatively more responsive to the reaction of balck shank in the explanation, and the disease between this after being ill the 5th～7 day of inoculation refers to change bigger.The result sees table 4 for details.
The disease of 5d refers to refer to mutation analysis with the disease of 7d after the different resistant variety inoculation of table 4 balck shank
| | 5d | 7d | 10d | Average disease refers to | Evaluation of resistance | Difference 1 | ||
| Coker Tobacco | 6.67 | 6.67 | 7.50 | 6.94 | |
0 | ||
| The COM cigarette | 4.17 | 4.17 | 13.33 | 7.22 | |
0 | ||
| Crust draws No. 1 | 25.00 | 25.83 | 30.83 | 27.22 | MR | 0.83 | ||
| Build ripple No. 1 | 0.00 | 0.83 | 3.33 | 1.39 | R | 0.83 | ||
| Peace is selected 79-1 | 5.00 | 5.83 | 10.83 | 7.22 | R | 0.83 | ||
| NC100 | 19.17 | 20.83 | 23.33 | 21.11 | R | 1.66 | ||
| OX414 | 5.00 | 6.67 | 12.50 | 8.06 | R | 1.67 | ||
| H4 | 20.00 | 21.67 | 35.83 | 25.83 | MR | 1.67 | ||
| H9 | 70.83 | 72.50 | 89.72 | 80.28 | S | 1.67 | ||
| 99F112 | 26.67 | 29.17 | 37.50 | 31.11 | MR | 2.5 | ||
| Land Huang | 2.50 | 5.00 | 8.33 | 5.28 | R | 2.5 | ||
| Draw No. 1 in Tianjin | 0.00 | 2.50 | 5.00 | 2.50 | R | 2.5 | ||
| MC1 | 32.50 | 35.83 | 50.00 | 39.44 | MR | 3.33 | ||
| Crust draws No. 2 | 10.00 | 13.33 | 15.83 | 13.06 | R | 3.33 | ||
| Carifoniavalle | 23.33 | 26.67 | 34.17 | 28.06 | MR | 3.34 | ||
| Pan Yuanhuang | 12.69 | 16.67 | 23.33 | 17.56 | R | 3.98 | ||
| H16 | 32.50 | 36.67 | 50.00 | 39.72 | MR | 4.17 | ||
| No. 7, platform cigarette | 20.00 | 24.17 | 25.83 | 23.33 | R | 4.17 | ||
| Fragrant No. 1 of cloud | 25.83 | 30.83 | 34.17 | 30.28 | MR | 5 | ||
| H6 | 34.17 | 39.17 | 50.00 | 41.11 | MR | 5 | ||
| 50926 | 60.83 | 66.67 | 71.67 | 66.39 | MS | 5.84 | ||
| H80A432 | 4.17 | 10.83 | 11.67 | 8.89 | R | 6.66 | ||
| The Xiao Bai muscle | 38.33 | 45.00 | 50.00 | 44.44 | MR | 6.67 | ||
| 9908 | 29.17 | 36.67 | 43.33 | 36.39 | MR | 7.5 |
Continuous table 4:
| 99K521 | 19.17 | 26.67 | 46.67 | 30.83 | MR | 7.5 |
| 2009 | 25.00 | 32.50 | 48.75 | 35.42 | MR | 7.5 |
| Is langold | 33.33 | 40.83 | 45.45 | 39.87 | MR | 7.5 |
| N.Syivestris | 75.00 | 82.50 | 85.00 | 80.83 | S | 7.5 |
| Hubei Province cigarette 1# | 26.67 | 35.00 | 42.50 | 34.72 | MR | 8.33 |
| Draw No. 3 in Tianjin | 7.50 | 15.83 | 22.50 | 15.28 | R | 8.33 |
| No. 5, platform cigarette | 38.33 | 46.67 | 56.67 | 47.22 | MR | 8.34 |
| 8022 | 24.17 | 33.33 | 43.33 | 33.61 | MR | 9.16 |
| H11 | 45.83 | 55.00 | 58.33 | 53.06 | MS | 9.17 |
| 2017 | 17.50 | 28.33 | 31.67 | 25.83 | MR | 10.83 |
| No. 11, Bath agate | 30.00 | 40.83 | 50.83 | 40.56 | MR | 10.83 |
| Huang Miao | 45.83 | 56.67 | 67.50 | 56.67 | MS | 10.84 |
| No. 12, Bath agate | 13.33 | 25.00 | 43.33 | 27.22 | MR | 11.67 |
| Cande1 | 57.50 | 70.00 | 77.50 | 68.33 | MS | 12.5 |
| H1 | 35.83 | 49.17 | 72.31 | 52.44 | MS | 13.34 |
| H15 | 43.33 | 60.00 | 75.00 | 59.44 | MS | 16.67 |
| YNBS1 | 49.17 | 67.50 | 69.17 | 61.94 | MS | 18.33 |
| H2 | 21.02 | 40.28 | 55.00 | 38.77 | MR | 19.26 |
| H13 | 38.52 | 60.00 | 68.33 | 55.62 | MS | 21.48 |
| H7 | 30.83 | 53.33 | 64.17 | 49.44 | MR | 22.5 |
| H12 | 24.17 | 46.67 | 56.67 | 42.50 | MR | 22.5 |
| H17 | 25.83 | 48.33 | 58.33 | 44.17 | MR | 22.5 |
| TN86 | 11.67 | 35.00 | 35.83 | 27.50 | MR | 23.33 |
| H10 | 40.00 | 64.17 | 72.50 | 58.89 | MS | 24.17 |
| H5 | 17.50 | 43.33 | 52.50 | 37.78 | MR | 25.83 |
| H8 | 45.83 | 75.00 | 77.50 | 66.11 | MS | 29.17 |
When referring to refer to subtract each other with 7 days disease with 10 days diseases: both diseases refer to that difference accounts for 64% of tested variety in the kind below 10, and wherein disease-resistant variety (R) has 13 parts, accounts for 100% of disease-resistant variety sum; In anti-kind (MR) have 11 parts, in accounting for 44% of anti-kind sum; Middle sense kind (MS) has 7 parts, feels 70% of kind sum in accounting for; Susceptible variety (S) has 1 part, accounts for 50% of susceptible variety sum.Difference accounts for 36% of tested variety in the kind more than 10, and anti-in being, middle sense and susceptible variety, wherein in anti-kind (MR) have 14 parts; In accounting for 56% of anti-kind sum; Middle sense kind (MS) has 3 parts, feels 30% of kind sum in accounting for; Susceptible variety (S) has 1 part, accounts for 50% of susceptible variety sum.
Whole disease-resistant varieties and 70% susceptible variety refer to difference below 10 the 7th～10 day disease, in ratio anti-and susceptible variety lower; And difference is in the kind more than 10, have 56% in anti-kind, and umber is also maximum, reaches 14 parts; Though there is 50% susceptible variety disease to refer to difference more than 10,1 part of susceptible variety is only arranged, umber is compared less.The result sees table 5 for details.
The disease of 7d refers to refer to mutation analysis with the disease of 10d after the different resistant variety inoculation of table 5 balck shank
| Variety name | 5d | 7d | 10d | Average disease refers to | Evaluation of resistance | Difference 2 |
| TN86 | 11.67 | 35.00 | 35.83 | 27.50 | MR | 0.83 |
| Coker Tobacco | 6.67 | 6.67 | 7.50 | 6.94 | R | 0.83 |
| H80A432 | 4.17 | 10.83 | 11.67 | 8.89 | R | 0.84 |
| No. 7, platform cigarette | 20.00 | 24.17 | 25.83 | 23.33 | R | 1.66 |
| YNBS1 | 49.17 | 67.50 | 69.17 | 61.94 | MS | 1.67 |
| Build ripple No. 1 | 0.00 | 0.83 | 3.33 | 1.39 | R | 2.50 |
| NC100 | 19.17 | 20.83 | 23.33 | 21.11 | R | 2.50 |
| Draw No. 1 in Tianjin | 0.00 | 2.50 | 5.00 | 2.50 | R | 2.50 |
| Crust draws No. 2 | 10.00 | 13.33 | 15.83 | 13.06 | R | 2.50 |
| N.Syivestris | 75.00 | 82.50 | 85.00 | 80.83 | S | 2.50 |
| H8 | 45.83 | 75.00 | 77.50 | 66.11 | MS | 2.50 |
| H11 | 45.83 | 55.00 | 58.33 | 53.06 | MS | 3.33 |
| Land Huang | 2.50 | 5.00 | 8.33 | 5.28 | R | 3.33 |
| Fragrant No. 1 of cloud | 25.83 | 30.83 | 34.17 | 30.28 | MR | 3.34 |
| 2017 | 17.50 | 28.33 | 31.67 | 25.83 | MR | 3.34 |
| Islangold | 33.33 | 40.83 | 45.45 | 39.87 | MR | 4.62 |
| Crust draws No. 1 | 25.00 | 25.83 | 30.83 | 27.22 | MR | 5.00 |
| Peace is selected 79-1 | 5.00 | 5.83 | 10.83 | 7.22 | R | 5.00 |
| 50926 | 60.83 | 66.67 | 71.67 | 66.39 | MS | 5.00 |
| The Xiao Bai muscle | 38.33 | 45.00 | 50.00 | 44.44 | MR | 5.00 |
| OX414 | 5.00 | 6.67 | 12.50 | 8.06 | R | 5.83 |
| Pan Yuanhuang | 12.69 | 16.67 | 23.33 | 17.56 | R | 6.66 |
| 9908 | 29.17 | 36.67 | 43.33 | 36.39 | MR | 6.66 |
| Draw No. 3 in Tianjin | 7.50 | 15.83 | 22.50 | 15.28 | R | 6.67 |
| Carifoniavalle | 23.33 | 26.67 | 34.17 | 28.06 | MR | 7.50 |
| Hubei Province cigarette 1# | 26.67 | 35.00 | 42.50 | 34.72 | MR | 7.50 |
| Cande1 | 57.50 | 70.00 | 77.50 | 68.33 | MS | 7.50 |
| 99F112 | 26.67 | 29.17 | 37.50 | 31.11 | MR | 8.33 |
| H13 | 38.52 | 60.00 | 68.33 | 55.62 | MS | 8.33 |
| H10 | 40.00 | 64.17 | 72.50 | 58.89 | MS | 8.33 |
| The COM cigarette | 4.17 | 4.17 | 13.33 | 7.22 | R | 9.16 |
| H5 | 17.50 | 43.33 | 52.50 | 37.78 | MR | 9.17 |
| No. 5, platform cigarette | 38.33 | 46.67 | 56.67 | 47.22 | MR | 10.00 |
| 8022 | 24.17 | 33.33 | 43.33 | 33.61 | MR | 10.00 |
| No. 11, Bath agate | 30.00 | 40.83 | 50.83 | 40.56 | MR | 10.00 |
| H12 | 24.17 | 46.67 | 56.67 | 42.50 | MR | 10.00 |
| H17 | 25.83 | 48.33 | 58.33 | 44.17 | MR | 10.00 |
Continuous table 5:
| H6 | 34.17 | 39.17 | 50.00 | 41.11 | MR | 10.83 |
| Huang Miao | 45.83 | 56.67 | 67.50 | 56.67 | MS | 10.83 |
| H7 | 30.83 | 53.33 | 64.17 | 49.44 | MR | 10.84 |
| H16 | 32.50 | 36.67 | 50.00 | 39.72 | MR | 13.33 |
| H4 | 20.00 | 21.67 | 35.83 | 25.83 | MR | 14.16 |
| MC1 | 32.50 | 35.83 | 50.00 | 39.44 | MR | 14.17 |
| H2 | 21.02 | 40.28 | 55.00 | 38.77 | MR | 14.72 |
| H15 | 43.33 | 60.00 | 75.00 | 59.44 | MS | 15.00 |
| 2009 | 25.00 | 32.50 | 48.75 | 35.42 | MR | 16.25 |
| H9 | 70.83 | 72.50 | 89.72 | 80.28 | S | 17.22 |
| No. 12, Bath agate | 13.33 | 25.00 | 43.33 | 27.22 | MR | 18.33 |
| 99K521 | 19.17 | 26.67 | 46.67 | 30.83 | MR | 20.00 |
| H1 | 35.83 | 49.17 | 72.31 | 52.44 | MS | 23.14 |
When referring to refer to subtract each other with 5 days disease with 10 days diseases: both diseases refer to that difference accounts for 64% of tested variety in the kind below 10, and wherein disease-resistant variety (R) has 13 parts, accounts for 100% of disease-resistant variety sum; In anti-kind (MR) have 15 parts, in accounting for 60% of anti-kind sum; Middle sense kind (MS) has 2 parts, feels 20% of kind sum in accounting for; Susceptible variety (S) has 2 parts, accounts for 100% of susceptible variety.Difference accounts for 36% of tested variety in the kind more than 10, and anti-in being, middle sense kind, wherein in anti-kind (MR) have 10 parts, in accounting for 40% of anti-kind sum; Middle sense kind (MS) has 8 parts, feels 80% of kind sum in accounting for.
Whole disease-resistant variety (R) and susceptible variety (S) refer to that the 5th～10 day disease difference is all below 10; And difference in more than 10 anti-and in the sense kind, 80% middle sense kind disease in this is identified refers to difference more than 10, the sense kind is relatively more responsive to the reaction of balck shank in the explanation, the disease between this after being ill the 5th～10 day of inoculation refers to change still bigger.The result sees table 6 for details.
The disease of 5d refers to refer to mutation analysis with the disease of 10d after the different resistant variety inoculation of table 6 balck shank
| | 5d | 7d | 10d | Average disease refers to | Evaluation of resistance | Difference 3 | ||
| Coker Tobacco | 6.67 | 6.67 | 7.50 | 6.94 | R | 0.27 | ||
| Build ripple No. 1 | 0.00 | 0.83 | 3.33 | 1.39 | R | 1.39 | ||
| NC100 | 19.17 | 20.83 | 23.33 | 21.11 | R | 1.94 | ||
| Crust draws No. 1 | 25.00 | 25.83 | 30.83 | 27.22 | MR | 2.22 | ||
| Peace is selected 79-1 | 5.00 | 5.83 | 10.83 | 7.22 | R | 2.22 | ||
| Draw No. 1 in Tianjin | 0.00 | 2.50 | 5.00 | 2.50 | R | 2.50 | ||
| Land Huang | 2.50 | 5.00 | 8.33 | 5.28 | R | 2.78 | ||
| The COM cigarette | 4.17 | 4.17 | 13.33 | 7.22 | R | 3.05 | ||
| Crust draws No. 2 | 10.00 | 13.33 | 15.83 | 13.06 | R | 3.06 |
Continuous table 6:
| OX414 | 5.00 | 6.67 | 12.50 | 8.06 | R | 3.06 |
| No. 7, platform cigarette | 20.00 | 24.17 | 25.83 | 23.33 | R | 3.33 |
| 99F112 | 26.67 | 29.17 | 37.50 | 31.11 | MR | 4.44 |
| Fragrant No. 1 of cloud | 25.83 | 30.83 | 34.17 | 30.28 | MR | 4.45 |
| H80A432 | 4.17 | 10.83 | 11.67 | 8.89 | R | 4.72 |
| Carifoniavalle | 23.33 | 26.67 | 34.17 | 28.06 | MR | 4.73 |
| Pan Yuanhuang | 12.69 | 16.67 | 23.33 | 17.56 | R | 4.87 |
| 50926 | 60.83 | 66.67 | 71.67 | 66.39 | MS | 5.56 |
| N.Syivestris | 75.00 | 82.50 | 85.00 | 80.83 | S | 5.83 |
| H4 | 20.00 | 21.67 | 35.83 | 25.83 | MR | 5.83 |
| The Xiao Bai muscle | 38.33 | 45.00 | 50.00 | 44.44 | MR | 6.11 |
| Islangold | 33.33 | 40.83 | 45.45 | 39.87 | MR | 6.54 |
| H6 | 34.17 | 39.17 | 50.00 | 41.11 | MR | 6.94 |
| MC1 | 32.50 | 35.83 | 50.00 | 39.44 | MR | 6.94 |
| 9908 | 29.17 | 36.67 | 43.33 | 36.39 | MR | 7.22 |
| H16 | 32.50 | 36.67 | 50.00 | 39.72 | MR | 7.22 |
| H11 | 45.83 | 55.00 | 58.33 | 53.06 | MS | 7.23 |
| Draw No. 3 in Tianjin | 7.50 | 15.83 | 22.50 | 15.28 | R | 7.78 |
| Hubei Province cigarette 1# | 26.67 | 35.00 | 42.50 | 34.72 | MR | 8.05 |
| 2017 | 17.50 | 28.33 | 31.67 | 25.83 | MR | 8.33 |
| No. 5, platform cigarette | 38.33 | 46.67 | 56.67 | 47.22 | MR | 8.89 |
| 8022 | 24.17 | 33.33 | 43.33 | 33.61 | MR | 9.44 |
| H9 | 70.83 | 72.50 | 89.72 | 80.28 | S | 9.45 |
| 2009 | 25.00 | 32.50 | 48.75 | 35.42 | MR | 10.42 |
| No. 11, Bath agate | 30.00 | 40.83 | 50.83 | 40.56 | MR | 10.56 |
| Cande1 | 57.50 | 70.00 | 77.50 | 68.33 | MS | 10.83 |
| Huang Miao | 45.83 | 56.67 | 67.50 | 56.67 | MS | 10.84 |
| 99K521 | 19.17 | 26.67 | 46.67 | 30.83 | MR | 11.66 |
| YNBS1 | 49.17 | 67.50 | 69.17 | 61.94 | MS | 12.77 |
| No. 12, Bath agate | 13.33 | 25.00 | 43.33 | 27.22 | MR | 13.89 |
| TN86 | 11.67 | 35.00 | 35.83 | 27.50 | MR | 15.83 |
| H15 | 43.33 | 60.00 | 75.00 | 59.44 | MS | 16.11 |
| H1 | 35.83 | 49.17 | 72.31 | 52.44 | MS | 16.61 |
| H13 | 38.52 | 60.00 | 68.33 | 55.62 | MS | 17.10 |
| H2 | 21.02 | 40.28 | 55.00 | 38.77 | MR | 17.75 |
| H12 | 24.17 | 46.67 | 56.67 | 42.50 | MR | 18.33 |
| H17 | 25.83 | 48.33 | 58.33 | 44.17 | MR | 18.34 |
| H7 | 30.83 | 53.33 | 64.17 | 49.44 | MR | 18.61 |
| H10 | 40.00 | 64.17 | 72.50 | 58.89 | MS | 18.89 |
| H8 | 45.83 | 75.00 | 77.50 | 66.11 | MS | 20.28 |
| H5 | 17.50 | 43.33 | 52.50 | 37.78 | MR | 20.28 |
Analysis-by-synthesis, three differences account for 48% of tested variety all at have 24 parts below 10, wherein disease-resistant variety (R) has 13 parts, in anti-kind (MR) have 8 parts, middle sense kind (MS) has 2 parts, susceptible variety (S) has 1 part; Three differences account for 18% of tested variety all at have 9 parts more than 10, and anti-in being, middle sense kind, wherein in anti-(MR) have 6 parts, middle sense kind (MS) has 3 parts.
What disease referred to that difference do not change when 7 days disease referred to refer to subtract each other with 5 days disease has 2 parts: Cokertobacco and COM cigarette, be disease-resistant variety (R), and disease refers to that it is H8 that difference changes maximum, is middle sense kind (MS); Disease referred to change when 10 days disease referred to refer to subtract each other with 7 days disease minimum has 2 parts: middle sense kind TN86, disease-resistant variety Coker tobacco.That change maximum is middle sense kind H1; When 10 days disease referred to refer to subtract each other with 5 days disease: it was to be still disease-resistant variety Coker tobacco that difference changes minimum, and it is middle sense kind H8 that difference changes maximum.The result as shown in Figure 1.
Claims (6)
1, a kind of sprout period testifying method of black shank resistance comprises the step of following order:
1. pathogen separates
Adding content in basal medium is the rifampin solution of 3000ug/ml, and it is standby to make the rifampin medium; Gather the typical ill tobacco stem of black shank symptom, get disease stem epidermis hypochondriasis and organize several piece, be cut into the fritter of long 1～2cm, place on the rifampin medium, behind cultivation, purifying, obtain the black shank mycelium, at ambient temperature, be stored in the sterile purified water standby;
2. the preparation of balck shank inoculum
With the black shank mycelium move into cultivate 5～7 days in the oat culture fluid after, change over to cultivate on the paddy medium and obtained the balck shank inoculum in 18～20 days; Described paddy medium is made by following method: the millet immersion was cleaned with clear water after 6 hours, wash in a pan shrivelled ghost grain to the greatest extent, boil and blow into half-open flower-shapedly to grain, cooling is dried in the air and is divided in the triangular flask of packing into to millet water content 50%～60% back, and 115 ℃ of following autoclavings 1 hour promptly.
3. design of sick garden and management
To the greenhouse disease garden, the cigarette seedling is randomized arrangement in sick garden with 45～55 days cigarette transplantation of seedlings to be identified of seedling age, establishes 3 repetitions, whenever repeats to plant cigarette 10 strains, and greenhouse management technology routinely manages; With little gold 1025 is susceptible contrast, and the big gold dollar of safflower is susceptible auxiliary contrast, and Venus 6007 resists contrast in being, reforming No. 3 is disease-resistant contrast;
4. bacterium paddy inoculation
After the cigarette transplantation of seedlings 5～7 days, will cultivate the basal part of stem that the inoculum that obtains is seeded to cigarette seedling to be measured through step (2), inoculum concentration is the 0.5g/ strain; After the inoculation immediately earthing and and the 15～20cm that pours water, kept 24～48 hours;
5. identify and classification
In the greenhouse disease garden, write down respectively once respectively at postvaccinal the 5th day, the 7th day and the 10th day that disease refers to, the mean value that refers to three diseases is as the resistance index of test kind; And then identification of species is carried out classification by national standard.
2, resistance authentication method according to claim 1 is characterized in that: the described basal medium of step (1) is an oat medium.
3, resistance authentication method according to claim 1 is characterized in that: the volume percent content of rifampin stoste is 1.3% in the described rifampin medium of step (1).
4, resistance authentication method according to claim 1 is characterized in that: the grain amount of packing in step (2) triangular flask is no more than 3/4 bottle.
5, resistance authentication method according to claim 1 is characterized in that: the described inoculation of step (4) is wound or the inoculation of non-wound.
6, resistance authentication method according to claim 5 is characterized in that: the described inoculation method of step (4) adopts not wound inoculation.
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Families Citing this family (17)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101491199B (en) * | 2009-03-09 | 2011-01-05 | 云南省烟草科学研究所 | Tobacco black shank resistance disease nursery identification method |
| CN101914454A (en) * | 2010-08-12 | 2010-12-15 | 云南省烟草农业科学研究院 | Simple and convenient separation method for phytophthora parasitica var nicotianae |
| CN101999287B (en) * | 2010-09-14 | 2012-01-04 | 云南省烟草农业科学研究院 | Thermostatic hydroponic identification method for resistance of tobacco black shank at seedling stage |
| CN102220437B (en) * | 2011-04-27 | 2014-04-23 | 贵州省烟草科学研究所 | Method for identifying viral disease resistance of tobacco through partial inoculation |
| CN102498878B (en) * | 2011-11-03 | 2014-04-09 | 云南省烟草农业科学研究院 | New method for identifying resistance in black shank quickly |
| CN102559516A (en) * | 2012-01-31 | 2012-07-11 | 云南省烟草农业科学研究院 | Tobacco black shank pathogen liquid-solid composite system culturing method and application of cultures |
| CN102523907A (en) * | 2012-02-10 | 2012-07-04 | 江西省农业科学院植物保护研究所 | Method for identifying resistance of asparagus to wilt disease in seedling stage |
| CN102598982B (en) * | 2012-03-20 | 2013-04-03 | 中国烟草中南农业试验站 | Method for testing cold resistance of tobacco in seedling stage |
| CN102835288A (en) * | 2012-08-28 | 2012-12-26 | 新疆农垦科学院 | Method for establishing disease garden to identify soil-borne diseases |
| CN103081678B (en) * | 2013-01-14 | 2014-01-08 | 中国农业科学院烟草研究所 | Rapid identification method of tobacco black shank resistance |
| CN104152533B (en) * | 2014-08-07 | 2016-06-29 | 贵州省烟草科学研究院 | One method growing tobacco seedling phase Rapid identification black shank fastness |
| CN104745672B (en) * | 2015-03-04 | 2017-06-23 | 河南省农业科学院烟草研究所 | A kind of method of Rapid identification tobacco black shank resistance |
| CN105075698B (en) * | 2015-09-25 | 2017-07-11 | 云南省烟草农业科学研究院 | A kind of authentication method of tobacco black shank resistance individual plant |
| CN105177104A (en) * | 2015-10-15 | 2015-12-23 | 中国烟草总公司郑州烟草研究院 | Method for identifying phytophthora parasitica var nicotianae physiological strains |
| CN106472141A (en) * | 2016-10-17 | 2017-03-08 | 云南省烟草农业科学研究院 | A kind of method of Artificial disease nursery Rapid identification tobacco black shank resistance |
| CN109392646A (en) * | 2018-11-23 | 2019-03-01 | 云南省烟草农业科学研究院 | A kind of investigation method of tobacco black shank resistance |
| CN113498722A (en) * | 2021-07-02 | 2021-10-15 | 中国农业科学院烟草研究所(中国烟草总公司青州烟草研究所) | Method for rapidly identifying resistance of tobacco black shank |
-
2006
- 2006-09-05 CN CNB2006100486585A patent/CN100446655C/en active Active
Non-Patent Citations (6)
| Title |
|---|
| 烟草黑胫病菌研究进展（二）. 马国胜，高智谋.烟草科技，第1期. 2004 |
| 烟草黑胫病菌研究进展（二）. 马国胜，高智谋.烟草科技，第1期. 2004 * |
| 烟草黑胫病菌致病性分化和烟草品种的抗病性差异. 梁元存等.植物保护学报，第30卷第2期. 2003 |
| 烟草黑胫病菌致病性分化和烟草品种的抗病性差异. 梁元存等.植物保护学报，第30卷第2期. 2003 * |
| 烤烟杂交组合比较试验. 卢秀萍等.云南农业大学学报，第18卷第1期. 2003 |
| 烤烟杂交组合比较试验. 卢秀萍等.云南农业大学学报，第18卷第1期. 2003 * |
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