CN104152533B - One method growing tobacco seedling phase Rapid identification black shank fastness - Google Patents
One method growing tobacco seedling phase Rapid identification black shank fastness Download PDFInfo
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- CN104152533B CN104152533B CN201410386031.5A CN201410386031A CN104152533B CN 104152533 B CN104152533 B CN 104152533B CN 201410386031 A CN201410386031 A CN 201410386031A CN 104152533 B CN104152533 B CN 104152533B
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Abstract
The invention discloses a method growing tobacco seedling phase Rapid identification black shank fastness, it is seeded in seedling culture hole plate by tobacco bred to be identified nursery, after seedling age reaches 35～45 days, carry out artificial vaccination, in the cavities of hole tray, imbed, for examination cigarette strain, the balck shank inoculation material 0.1～1g cultivated in advance to all, within 3～10 days, investigate severity Scaling after inoculation respectively, calculate disease index.The present invention adopts the authentication method that seedling is not transplanted/transplanted, and does not artificially create wound, more efficient and convenient, through the checking synchronization with the inventive method of conventional field Disease garden identification for many years, qualification result of the present invention is suitable with field test, and between year, difference is little, qualification time is short, and workload is little.
Description
Technical field
The invention belongs to disease resistance of plant identification technology field, be specifically related to tobacco black shank resistance authentication method.
Background technology
Black shank is one of most destructive disease on tobacco leaf production, is also endanger one of the most serious disease in World tobacco production simultaneously, and this disease is respectively produced cigarette in China and economized the generation all having in various degree, and harm is comparatively general and serious.Selecting the anti-balck shank kind of Nicotiana tabacum L. is solve the important channel of this disease.
At present, existing tobacco bred method of resistance identification mainly has field balck shank garden identification method and sprout period authenticating method.Balck shank garden, field identification method is owing to by soil fertility, season, annual identification of species quantity is very limited, and the varietal resistance identified between the time has certain difference；And existing sprout period authenticating method will be transplanted in flowerpot, indoor sick garden after nursery or be transplanted on new dish, shortcoming is to transplant inoculation after slow Seedling, and the time is long, and workload is big.
Summary of the invention
The technical problem to be solved in the present invention is: for problems of the prior art, the invention provides the authentication method that a kind of seedling is not transplanted/transplanted, artificially do not create wound, more efficient and convenient, through the synchronization contrast verification of field Disease garden identification for many years with the inventive method, qualification result of the present invention is suitable with field test, and between year, difference is little, qualification time is short, and workload is few.
The technical solution used in the present invention: seedling Rapid identification method carries out in greenhouse, temperature environment 25 DEG C.Each tobacco bred to be identified is seeded into nursery in 54 hole seedling culture hole plates, each kind/it is 1 dish, seedling management measure is with conventional leaf tobacco production.After seedling age reaches 35～45 days, (do not need leaf-cutting), carry out artificial vaccination.In the cavities of hole tray, imbed, for examination cigarette strain, the balck shank inoculation material 0.1～1g cultivated in advance to all.Within 3～10 days, investigate severity Scaling after inoculation respectively, calculate disease index.
The making of inoculation material: by ripe for Semen setariae decocting in water to 6 point, pulls out and is put on gauze and spreads out, dewatering 10 minutes.Load in 250ml round plastic tissue culture bottle, be filled to 2/3 place of bottle.With 121 DEG C of sterilizings of high-pressure steam sterilizing pot 1 hour.After being cooled to room temperature, on the balck shank mycelium inoculation on oat medium of separator well to Semen setariae, cultivating 15 days in incubator at 28 DEG C, mycelia is covered with.
Record method performs national standard " tobacco disease insect pest classification and investigation method (GB/T23222-2008) ".Disease index (D) computing formula:
D=∑ (diseased plant numbers at different levels × this disease progression) × 100/(investigates total strain number × superlative degree number)
Disease severity grade scale performs by standard GB/T/T23224-2008, and Partition in resistance is 6 ranks: high resistance or immunity (I): disease refers to be 0；Disease-resistant (R): disease-resistant (R)；In anti-(MR): disease refers to be 20.1～40；Middle sense (MS): disease refers to be 40.1～60；Sense (S): disease refers to be 60.1～80；High sense (HS): disease refers to be 80.1～100.
The beneficial effect that the present invention reaches:
The present invention adopts the authentication method that seedling is not transplanted/transplanted, and does not artificially create wound, more efficient and convenient.Through the checking synchronization with the inventive method of conventional field Disease garden identification for many years, qualification result of the present invention is suitable with field test, and between year, difference is little.Concrete manifestation is as follows:
(1) qualification cycle is shorter, and namely seedling not leaf-cutting in 35～45 days can be used for identifying, is only whole qualification cycle 45～55 days, more shorter than the qualification cycle of other authentication method existing, illustrates that the method for the present invention is more efficient；
(2) breeding method is different, and other method all adopts floating seedlings, identifies or is transplanted on new building and identify after transplanting.This method adopts macropore hole tray, has sufficient space Direct Identification on former dish, and dish does not soak in nutritional solution, is more suitable for the inoculation of solid vaccination cause of disease；
(3) making of cause of disease inoculation material is different.It is adopt the millet with shell to make that conventional bacterium paddy makes, and the Semen setariae after the employing threshing of this method makes, more convenient inoculation, and is conducive to morbidity.
(4) Semen setariae used by balck shank inoculation material through decocting in water, drain, sterilize, be filled in rectangle cell body and carry out uniform piecemeal, directly take by block so in use, so can improve test rate further, improve test specification degree.
Detailed description of the invention
Embodiment 1
1, materials and methods
1.1, test material
Guizhou Province's tobacco bred/and it is 10 parts of flue-cured tobacco materials, check variety innovation 3, Venus 6007, little gold 1025.
1.2, test method
1.2.1, conventional Disease garden identification method
Field Disease garden identification is tested in balck shank qualification garden in Guizhou Province Tabacco Science and Technology Institute flue-cured tobacco cultivars and is carried out.Test sets 4 repetitions, often repeats 1 row, plants cigarette 10 strain.Adopting floating seedlings method, method for culturing seedlings produces with conventional.Leaf-cutting twice, after cultivating into strong sprout, transplants the first tenday period of a month in May, density 1 × 0.6m2, mu executes purity nitrogen 6 kilograms.Other same Production of Large Fields of cultivation management measure.If reform No. 3 for disease-resistant comparison, Venus 6007 be in anti-comparison, little gold 1025 is susceptible comparison.Field test, in transplanting latter 20 days, carries out artificial vaccination to all for examination cigarette strain, imbeds the balck shank inoculation material 3g cultivated in advance at cigarette strain root.Within 20 days, 40 days, 60 days, investigate severity Scaling after inoculation respectively, take three investigation mean value calculation disease indexs.
1.2.2, the inventive method
Seedling Rapid identification method carries out in greenhouse, temperature environment 25 DEG C.Each tobacco bred to be identified is seeded into nursery in 54 hole seedling culture hole plates, each kind/it is 1 dish.Seedling management measure is with conventional leaf tobacco production.After seedling age reaches 40 days, (do not need leaf-cutting), carry out artificial vaccination.In the cavities of hole tray, imbed, for examination cigarette strain, the balck shank inoculation material 0.5g cultivated in advance to all.Within 3 days, 7 days, 10 days, investigate severity Scaling after inoculation respectively, take three investigation mean value calculation disease indexs.
1.2.3, the making of inoculation material
By ripe for Semen setariae decocting in water to 6 point, pull out and be put on gauze and spread out, dewatering 10 minutes.Load in 250ml round plastic tissue culture bottle, be filled to 2/3 place of bottle.With 121 DEG C of sterilizings of high-pressure steam sterilizing pot 1 hour.After being cooled to room temperature, Semen setariae after sterilizing is uniformly filled in rectangular channel body, Semen setariae in 0.1～1g uniform rectangular cell body is cut into small pieces by weight, by on the balck shank mycelium inoculation on oat medium of separator well to Semen setariae, incubator is cultivated 15 days at 28 DEG C, mycelia is covered with, and takes by block during use.
1.2.4 investigation method and Partition in resistance
Record method performs national standard " tobacco disease insect pest classification and investigation method (GB/T23222-2008) ".
Disease index (D) computing formula:
D=∑ (diseased plant numbers at different levels × this disease progression) × 100/(investigates total strain number × superlative degree number)
Partition in resistance performs by standard GB/T/T23224-2008, is divided into 6 ranks:
Table 1: national standard Partition in resistance table
Table 2: conventional Disease garden identification and seedling Rapid identification result
Synchronization proving and comparisom by the inventive method and conventional Disease garden identification method, be can be seen that by the result of the test of table 2, the qualification result of two kinds of methods is basically identical to the judgement of varietal resistance, the effectiveness of the inventive method is described, and the qualification cycle of the method that the present invention adopts is shorter, workload is less, is the method for a kind of more superior Nicotiana tabacum L. Rapid identification black shank fastness in little seedling stage.
Claims (1)
1. a method growing tobacco seedling phase Rapid identification black shank fastness, it is characterized in that: tobacco bred to be identified is seeded into nursery in 54 hole seedling culture hole plates, each kind/it is 1 dish, seedling management measure is with conventional leaf tobacco production, after seedling age reaches 40 days, does not need transplanting, leaf-cutting, direct labor inoculates, in the cavities of hole tray, imbed, for examination cigarette strain, the balck shank inoculation material 0.5g cultivated in advance to all, within 3 days, 7 days and 10 days, investigate severity Scaling after inoculation respectively, calculate disease index；The manufacture method of the balck shank inoculation material cultivated in advance is for by ripe for Semen setariae decocting in water to 6 point, pull out and be put on gauze and spread out, dewatering 10 minutes, load in 250ml round plastic tissue culture bottle, be filled to 2/3 place of bottle, with 121 DEG C of sterilizings of high-pressure steam sterilizing pot 1 hour, after being cooled to room temperature, on the balck shank mycelium inoculation on oat medium of separator well to Semen setariae, cultivating 15 days in incubator at 28 DEG C, mycelia covers with to obtain balck shank inoculation material；Before inoculating balck shank mycelia on Semen setariae, being uniformly filled in rectangular channel body by the Semen setariae after sterilizing, the Semen setariae in 0.1～1g uniform rectangular cell body is cut into small pieces by weight, and balck shank mycelia is uniformly seeded on the Semen setariae material of bulk.
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| 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 |
| CN105567784A (en) * | 2016-02-05 | 2016-05-11 | 云南省烟草农业科学研究院 | Method for rapidly and accurately identifying brown spot resistance of tobacco variety |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1918959A (en) * | 2006-09-05 | 2007-02-28 | 云南省烟草科学研究所 | Sprout period authenticating method for tobacco mosaic resistantance |
| CN1922965A (en) * | 2006-09-05 | 2007-03-07 | 云南省烟草科学研究所 | Sprout period authenticating method for tobacco black shank fastness |
| CN102220437A (en) * | 2011-04-27 | 2011-10-19 | 贵州省烟草科学研究所 | Method for identifying viral disease resistance of tobacco through partial inoculation |
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Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1918959A (en) * | 2006-09-05 | 2007-02-28 | 云南省烟草科学研究所 | Sprout period authenticating method for tobacco mosaic resistantance |
| CN1922965A (en) * | 2006-09-05 | 2007-03-07 | 云南省烟草科学研究所 | Sprout period authenticating method for tobacco black shank fastness |
| CN102220437A (en) * | 2011-04-27 | 2011-10-19 | 贵州省烟草科学研究所 | Method for identifying viral disease resistance of tobacco through partial inoculation |
Non-Patent Citations (3)
| Title |
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| 烟草对黑胫病的抗性的新鉴定法;大桥雄司;《中国烟草》;19810430(第4期);第47-48页 * |
| 烟草种质资源苗期黑胫病抗性鉴定研究;于海芹等;《中国农业科技导报》;20080430;第10卷(第4期);第70-75页 * |
| 烟草黑胫病及其抗性鉴定方法研究进展;孙计平;《河南农业科学》;20110831;第40卷(第8期);第36-39页 * |
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