CN106350574A - Appraisal method for indoor inoculation of disease resistance in resistance of tobacco to black shank - Google Patents
Appraisal method for indoor inoculation of disease resistance in resistance of tobacco to black shank Download PDFInfo
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- CN106350574A CN106350574A CN201610764762.8A CN201610764762A CN106350574A CN 106350574 A CN106350574 A CN 106350574A CN 201610764762 A CN201610764762 A CN 201610764762A CN 106350574 A CN106350574 A CN 106350574A
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- balck shank
- tobacco
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/02—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving viable microorganisms
- C12Q1/18—Testing for antimicrobial activity of a material
Abstract
The invention discloses a method for indoor inoculation of disease resistance in resistance of tobacco to black shank, comprising the steps of: step 1: tobacco cultivars to be appraised, tobacco cultivars known to be resistant to black shank, as well as tobacco cultivars known to affected with black shank are planted in tobacco fields; step 2: when the tobacco grow true leaves, they are transplanted indoors. After the survival of tobacco seedlings, all tobacco is inoculated with Phytophthora parasitica var.nicotiana to induce the disease; step 3: the disease resistance of each cultivar against black shank disease is detected. The method for indoor inoculation of disease resistance in resistance of tobacco to black shank can effectively appraise the disease resistance of tobacco cultivars.
Description
Technical field
The present invention relates to the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L..
Background technology
Black shank is one of most destructive disease on tobacco leaf production, and also known as Nicotiana tabacum L. epidemic disease, tobacco grower is referred to as " black bar
Crazy ", " black root ", " Aconitum carmichjaelii Debx. disease ".The each main product cigarette district of China all has different degrees of generation, and wherein Anhui, Shandong, Henan Province are to go through
Severe disease area in history；The Main Tobacco-growing Regions In Souths such as Yunnan, Guizhou, Sichuan, Hunan, Guangdong, Guangxi, Fujian occur also fairly common and many
Mix generation with tobacco bacterial wilt, therefore harm is even more serious.Generally change kind of disease-resistant variety come the integrated control state of an illness by taking.As
The disease resistance of the anti-balck shank of what identification tobacco bred is strong and weak, and the tobacco bred selecting anti-balck shank is particularly important.
Content of the invention
The present invention proposes a strong and weak indoor inoculation authentication method of disease resistance growing tobacco anti-balck shank, can be effective
The disease resistance of identification tobacco bred is strong and weak.
The technical scheme is that and be achieved in that:
The strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L., comprises the following steps:
Step one, vega plant tobacco bred to be identified, known anti-balck shank tobacco bred, known in anti-balck shank cigarette
Grass product kind and known sense balck shank tobacco bred；
Step 2, when tobacco seedlings grow true leaf, transplants to interior, after tobacco seedlings survive, with inoculating the bacterium of balck shank strain
Paddy inoculates all Nicotiana tabacum L.s, induces disease；
Step 3, the disease resistance of the detection anti-balck shank of each kind is strong and weak.
Further, in step 2 when tobacco seedlings length is to 5-6 piece true leaf, transplant to interior, every kind plants 10 plants -20 plants,
After tobacco seedlings survive, press the inoculation of 0.25g-0.50g bacterium paddy amount for every plant, be placed in temperature 26-30 DEG C, relative temperature 75%-85%
Under the conditions of induce disease.
Further, also planted on vega in step one locality anti-balck shank tobacco bred, locality in resist black shin
The sense balck shank tobacco bred of sick tobacco bred and locality is as auxiliary comparison.
Preferably, described known anti-balck shank tobacco bred is " innovation No. three ", described known in anti-balck shank Nicotiana tabacum L. product
Planting is " Venus 6007 ", and described known sense balck shank tobacco bred is " little gold 1025 ".
Further, the preparation method of described bacterium paddy is as follows: millet is boiled and blooms to 1/3-2/3 grain, pull out, load triangle
Bottle, autoclaving 1-2h, cultured balck shank strain is inoculated into after sterilizing on the millet of cooling, be subsequently placed in 28 DEG C～
Culture 15d～20d at 30 DEG C.
Further, pass through in step 3 to detect the sickness rate of each kind and/or disease index and/or attacked trees rate and/or
There are aphid (worm) strain rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate to detect each product
The disease resistance planting anti-balck shank is strong and weak.In their early stage, contain phase and latter stage respectively detection 1 time.
Further, sickness rate=(morbidity strain number/investigation total strain number) * 100%；Disease index=[σ (diseased plants at different levels or leaf
Number this sick level value of *)/(investigation total strain number or the highest value of number of sheets *)] * 100；Attacked trees rate=(the total strain of killed strain number/investigation
Number) * 100%；There is aphid (worm) strain rate=[having aphid (worm) strain number/investigation total strain number] * 100%；Aphid volume index=[σ (leaves at different levels
Number this class value of *)/(investigation total strain number or the highest value of number of sheets *)] * 100；Average individual plant aphid amount=total aphid amount/total strain number；Hundred
Strain worm amount=(total worm amount/total strain number) * 100；There is ovum strain rate=(having ovum strain number/investigation total strain number) * 100%；Balck shank disease
Severity gradation is as follows: 0 grade: Herb is anosis；1 grade: stem's scab is less than 1/3rd of stem girth, or 1/3rd with inferior lobe
Piece is wilting；3 grades: stem's scab is around stem girth 1/1st to two/3rd, or 1/1st to two/3rd blade slightly withers
Wither, or scab in bottom minority blade；5 grades: stem's scab exceedes 1/2nd of stem girth, but not entirely around stem girth, or
2/to three/2nd blades are wilting；7 grades: stem's scab is entirely around stem girth, or 2/3rds is wilting with blade；9
Level: diseased plant is substantially withered.
The invention has the benefit that the strong and weak indoor inoculation identification side of the disease resistance of the anti-balck shank of the Nicotiana tabacum L. of the present invention
Method, can effectively identify that the disease resistance of tobacco bred is strong and weak.The effect of key can be played in terms of vega Nicotiana tabacum L. seed selection.
Specific embodiment
For more fully understanding the present invention, below by following examples, the present invention is made specifically to illustrate further, but not
Can be regarded as limitation of the invention, some made according to foregoing invention content for those skilled in the art are nonessential
Improve and adjustment, be also considered as being within the scope of the present invention.
Embodiment 1
The strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L., comprises the following steps:
Step one, vega plant tobacco bred to be identified, known anti-balck shank tobacco bred, known in anti-balck shank cigarette
Grass product kind and known sense balck shank tobacco bred, described known anti-balck shank tobacco bred is " innovation three ", described known
In anti-balck shank tobacco bred be " Venus 6007 ", described known sense balck shank tobacco bred be " little gold 1025 "；
Step 2, when tobacco seedlings length is to 5 true leaves, transplants to indoor flowerpot, a diameter of 12cm of flowerpot, 2 plants of every basin kind,
Every kind plants 10 plants, after tobacco seedlings survive, presses the inoculation of 0.25g bacterium paddy amount for every plant, is placed in 26 DEG C of temperature, relative temperature 75%
Under the conditions of induce disease, the preparation method of described bacterium paddy is as follows: millet is boiled and blooms to 1/3 grain, pull out, load triangular flask,
Autoclaving 1h, cultured balck shank strain is inoculated on the millet of cooling after sterilizing, is subsequently placed in culture at 28 DEG C
20d；
Step 3, by detecting the sickness rate of each kind and/or disease index and/or attacked trees rate and/or having aphid (worm)
Strain rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate detect each kind resist black shin
The disease resistance of disease is strong and weak, in their early stage, contains phase and latter stage respectively detection 1 time, sickness rate=(morbidity strain number/investigation total strain number) *
100%；Disease index=[σ (diseased plants at different levels or this sick level value of number of sheets *)/(investigation total strain number or the highest value of number of sheets *)] *
100；Attacked trees rate=(killed strain number/investigation total strain number) * 100%；There is aphid (worm) strain rate=[have aphid (worm) strain number/investigation total
Strain number] * 100%；Aphid volume index=[σ (this class value of numbers of sheets * at different levels)/(investigation total strain number or the highest value of number of sheets *)] *
100；Average individual plant aphid amount=total aphid amount/total strain number；Hundred plants of worm amount=(total worm amount/total strain numbers) * 100；There is ovum strain rate=(have ovum
Strain number/investigation total strain number) * 100%；The classification of balck shank disease severity is as follows: 0 grade: Herb is anosis；1 grade: stem's scab does not surpass
Cross 1/3rd of stem girth, or 1/3rd is wilting with lower blade；3 grades: stem scab around stem girth 1 to two/3rd/
One, or 1/1st to two/3rd blade is slightly wilting, or scab in bottom minority blade；5 grades: stem's scab exceedes stem
/ 2nd enclosing, but not entirely around stem girth, or 2/1sts to three/2nd blades are wilting；7 grades: the whole ring of stem's scab
Around stem girth, or 2/3rds is wilting with blade；9 grades: diseased plant is substantially withered.
Embodiment 2
The strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L., comprises the following steps:
Step one, vega plant tobacco bred to be identified, known anti-balck shank tobacco bred, known in anti-balck shank cigarette
Grass product kind, known sense balck shank tobacco bred, local anti-balck shank tobacco bred, in locality anti-balck shank tobacco bred with
And the sense balck shank tobacco bred of locality, described known anti-balck shank tobacco bred is " innovation No. three ", described known in anti-black
Shin disease tobacco bred is " Venus 6007 ", and described known sense balck shank tobacco bred is " little gold 1025 "；
Step 2, when tobacco seedlings length is to 6 true leaves, transplants to interior, and every kind plants 20 plants, after tobacco seedlings survive, often
The inoculation of 0.50g bacterium paddy amount is pressed in strain, is placed in 30 DEG C of temperature, induces disease, the preparation of described bacterium paddy under conditions of relative temperature 85%
Method is as follows: millet is boiled and blooms to 2/3 grain, pull out, load triangular flask, autoclaving 2h, by cultured balck shank
Strain is inoculated on the millet of cooling after sterilizing, is subsequently placed at 30 DEG C culture 15d；
Step 3, by detecting the sickness rate of each kind and/or disease index and/or attacked trees rate and/or having aphid (worm)
Strain rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate detect each kind resist black shin
The disease resistance of disease is strong and weak, in their early stage, contains phase and latter stage respectively detection 1 time, sickness rate=(morbidity strain number/investigation total strain number) *
100%；Disease index=[σ (diseased plants at different levels or this sick level value of number of sheets *)/(investigation total strain number or the highest value of number of sheets *)] *
100；Attacked trees rate=(killed strain number/investigation total strain number) * 100%；There is aphid (worm) strain rate=[have aphid (worm) strain number/investigation total
Strain number] * 100%；Aphid volume index=[σ (this class value of numbers of sheets * at different levels)/(investigation total strain number or the highest value of number of sheets *)] *
100；Average individual plant aphid amount=total aphid amount/total strain number；Hundred plants of worm amount=(total worm amount/total strain numbers) * 100；There is ovum strain rate=(have ovum
Strain number/investigation total strain number) * 100%；The classification of balck shank disease severity is as follows: 0 grade: Herb is anosis；1 grade: stem's scab does not surpass
Cross 1/3rd of stem girth, or 1/3rd is wilting with lower blade；3 grades: stem scab around stem girth 1 to two/3rd/
One, or 1/1st to two/3rd blade is slightly wilting, or scab in bottom minority blade；5 grades: stem's scab exceedes stem
/ 2nd enclosing, but not entirely around stem girth, or 2/1sts to three/2nd blades are wilting；7 grades: the whole ring of stem's scab
Around stem girth, or 2/3rds is wilting with blade；9 grades: diseased plant is substantially withered.
Embodiment 3
The strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L., comprises the following steps:
Step one, vega plant tobacco bred to be identified, known anti-balck shank tobacco bred, known in anti-balck shank cigarette
Grass product kind and known sense balck shank tobacco bred, described known anti-balck shank tobacco bred is " innovation three ", described known
In anti-balck shank tobacco bred be " Venus 6007 ", described known sense balck shank tobacco bred be " little gold 1025 "；
Step 2, when tobacco seedlings length is to 5 true leaves, transplants to interior, and every kind plants 15 plants, after tobacco seedlings survive, often
The inoculation of 0.35g bacterium paddy amount is pressed in strain, is placed in 28 DEG C of temperature, induces disease, the preparation of described bacterium paddy under conditions of relative temperature 80%
Method is as follows: millet is boiled and blooms to 1/2 grain, pull out, load triangular flask, autoclaving 1.5h, by cultured black shin
Pathogenic bacteria kind is inoculated on the millet of cooling after sterilizing, is subsequently placed at 29 DEG C culture 18d；
Step 3, by detecting the sickness rate of each kind and/or disease index and/or attacked trees rate and/or having aphid (worm)
Strain rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate detect each kind resist black shin
The disease resistance of disease is strong and weak, in their early stage, contains phase and latter stage respectively detection 1 time, sickness rate=(morbidity strain number/investigation total strain number) *
100%；Disease index=[σ (diseased plants at different levels or this sick level value of number of sheets *)/(investigation total strain number or the highest value of number of sheets *)] *
100；Attacked trees rate=(killed strain number/investigation total strain number) * 100%；There is aphid (worm) strain rate=[have aphid (worm) strain number/investigation total
Strain number] * 100%；Aphid volume index=[σ (this class value of numbers of sheets * at different levels)/(investigation total strain number or the highest value of number of sheets *)] *
100；Average individual plant aphid amount=total aphid amount/total strain number；Hundred plants of worm amount=(total worm amount/total strain numbers) * 100；There is ovum strain rate=(have ovum
Strain number/investigation total strain number) * 100%；The classification of balck shank disease severity is as follows: 0 grade: Herb is anosis；1 grade: stem's scab does not surpass
Cross 1/3rd of stem girth, or 1/3rd is wilting with lower blade；3 grades: stem scab around stem girth 1 to two/3rd/
One, or 1/1st to two/3rd blade is slightly wilting, or scab in bottom minority blade；5 grades: stem's scab exceedes stem
/ 2nd enclosing, but not entirely around stem girth, or 2/1sts to three/2nd blades are wilting；7 grades: the whole ring of stem's scab
Around stem girth, or 2/3rds is wilting with blade；9 grades: diseased plant is substantially withered.
The sickness rate of tobacco bred to be identified and/or disease index and/or attacked trees rate and/or have aphid (worm) strain rate and/
Or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate and known anti-balck shank tobacco bred
And/or the sickness rate of anti-balck shank tobacco bred of locality and/or disease index and/or attacked trees rate and/or have aphid (worm) strain
Rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have the same or like kind of ovum strain rate
For anti-balck shank kind；
The sickness rate of tobacco bred to be identified and/or disease index and/or attacked trees rate and/or have aphid (worm) strain rate and/
Or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate with known in anti-balck shank tobacco bred
And/or the sickness rate of anti-balck shank tobacco bred and/or disease index and/or attacked trees rate and/or have aphid (worm) in locality
Strain rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have the same or like product of ovum strain rate
Anti- balck shank kind in kind being；
The sickness rate of tobacco bred to be identified and/or disease index and/or attacked trees rate and/or have aphid (worm) strain rate and/
Or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate with known sense balck shank tobacco bred
And/or the sickness rate of sense balck shank tobacco bred of locality and/or disease index and/or attacked trees rate and/or have aphid (worm) strain
Rate and/or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have the same or like kind of ovum strain rate
For feeling balck shank kind.
The foregoing is only presently preferred embodiments of the present invention, not in order to limit the present invention, all essences in the present invention
Within god and principle, any modification, equivalent substitution and improvement made etc., should be included within the scope of the present invention.
Claims (8)
1. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. is it is characterised in that comprise the following steps:
Step one, vega plant tobacco bred to be identified, known anti-balck shank tobacco bred, known in anti-balck shank Nicotiana tabacum L. product
Plant and known sense balck shank tobacco bred；
Step 2, when tobacco seedlings grow true leaf, transplants to interior, after tobacco seedlings survive, is connect with the bacterium paddy inoculating balck shank strain
Plant all Nicotiana tabacum L.s, induce disease；
Step 3, the disease resistance of the detection anti-balck shank of each kind is strong and weak.
2. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 1 it is characterised in that:
In step 2 when tobacco seedlings length is to 5-6 piece true leaf, transplant to interior, every kind plants 10 plants -20 plants, after tobacco seedlings survive, often
The inoculation of 0.25g-0.50g bacterium paddy amount is pressed in strain, is placed in temperature 26-30 DEG C, induces disease under conditions of relative temperature 75%-85%.
3. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 1 it is characterised in that:
Also planted on vega in step one locality anti-balck shank tobacco bred, locality in anti-balck shank tobacco bred and work as
The sense balck shank tobacco bred on ground is as auxiliary comparison.
4. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 1 it is characterised in that:
Described known anti-balck shank tobacco bred is " innovation No. three ", described known in anti-balck shank tobacco bred be " Venus 6007 ",
Described known sense balck shank tobacco bred is " little gold 1025 ".
5. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 1 it is characterised in that:
The preparation method of described bacterium paddy is as follows: millet is boiled and blooms to 1/3-2/3 grain, pull out, load triangular flask, autoclaving 1-
2h, cultured balck shank strain is inoculated on the millet of cooling after sterilizing, is subsequently placed in culture 15d at 28 DEG C～30 DEG C
～20d.
6. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 1 it is characterised in that:
Pass through in step 3 to detect the sickness rate of each kind and/or disease index and/or attacked trees rate and/or have aphid (worm) strain rate and/
Or aphid volume index and/or average individual plant aphid amount and/or hundred plants of worm amounts and/or have ovum strain rate to detect the disease-resistant of the anti-balck shank of each kind
Property strong and weak.
7. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 6 it is characterised in that:
In their early stage, contain phase and latter stage respectively detection 1 time.
8. the strong and weak indoor inoculation authentication method of the disease resistance of the anti-balck shank of Nicotiana tabacum L. as claimed in claim 6 it is characterised in that:
Sickness rate=(morbidity strain number/investigation total strain number) * 100%；Disease index=[σ (diseased plants at different levels or this sick level value of number of sheets *)/
(investigation total strain number or the highest value of number of sheets *)] * 100；Attacked trees rate=(killed strain number/investigation total strain number) * 100%；There is aphid
(worm) strain rate=[having aphid (worm) strain number/investigation total strain number] * 100%；Aphid volume index=[σ (this class value of numbers of sheets * at different levels)/
(investigation total strain number or the highest value of number of sheets *)] * 100；Average individual plant aphid amount=total aphid amount/total strain number；Hundred plants of worm amount=(total worms
Amount/total strain number) * 100；There is ovum strain rate=(having ovum strain number/investigation total strain number) * 100%；Balck shank disease severity is classified such as
Under: 0 grade: Herb is anosis；1 grade: stem's scab is less than 1/3rd of stem girth, or 1/3rd is wilting with lower blade；3 grades:
Stem's scab is around stem girth 1/1st to two/3rd, or 1/1st to two/3rd blade is slightly wilting, or bottom is few
Scab in number blade；5 grades: stem's scab exceedes 1/2nd of stem girth, but not entirely around stem girth, or 1 to three/2nd
/ bis- blades are wilting；7 grades: stem's scab is entirely around stem girth, or 2/3rds is wilting with blade；9 grades: diseased plant is substantially withered
Extremely.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
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| CN201610764762.8A CN106350574A (en) | 2016-08-30 | 2016-08-30 | Appraisal method for indoor inoculation of disease resistance in resistance of tobacco to black shank |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111727784A (en) * | 2020-07-27 | 2020-10-02 | 贵州省烟草科学研究院 | Method for forecasting tobacco black shank initial period |
-
2016
- 2016-08-30 CN CN201610764762.8A patent/CN106350574A/en active Pending
Non-Patent Citations (2)
| Title |
|---|
| 于海芹等: "烟草品种黑胫病抗性鉴定及不同接种方法间的相关性分析", 《内蒙古农业科技》 * |
| 孙计平等: "烟草黑胫病及其抗性鉴定方法研究进展", 《河南农业科学》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111727784A (en) * | 2020-07-27 | 2020-10-02 | 贵州省烟草科学研究院 | Method for forecasting tobacco black shank initial period |
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