CN109220786A - A method of screening hypersecretion profile material from tobacco mutant body - Google Patents
A method of screening hypersecretion profile material from tobacco mutant body Download PDFInfo
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- CN109220786A CN109220786A CN201811374489.3A CN201811374489A CN109220786A CN 109220786 A CN109220786 A CN 109220786A CN 201811374489 A CN201811374489 A CN 201811374489A CN 109220786 A CN109220786 A CN 109220786A
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- screening
- hypersecretion
- tobacco
- identification
- seed
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/06—Processes for producing mutations, e.g. treatment with chemicals or with radiation
Abstract
The method that the invention discloses a kind of to screen hypersecretion profile material from tobacco mutant body library, comprising the following steps: step 1, seed treatment；Step 2, the preliminary screening of mutagenized populations；Step 3, single plant screening and identification；Step 4, repetitive identified (identification of F2 generation)；Step 5, ore grade indexes.The present invention is at identification initial stage, utilize the correlation of trichome density and type and blade face secretion, first carry out the screening of the density and type of intuitive blade face glandular hairs, carry out the identification of blade face secretion and the screening of hypersecretion profile material on this basis again, the selection result is accurate and reliable, 90% or more mutant plants have just been eliminated in Seedling Stage, greatly reduce the plantation amount of crop field material, reduce land used, the expenses such as recruitment, because of the reduction of planting material, decrease the quantity of maturity period secretion identification, great amount of cost is saved, shorten screening time, substantially increase screening efficiency.
Description
Technical field
The present invention relates to field of biotechnology, and in particular to a kind of side that hypersecretion profile material is screened from tobacco mutant body
Method.
Background technique
The quality and resistance of tobacco blade face secretion and tobacco leaf are closely related.Secretion content high kind cigarette strain in blade face is anti-
Property is strong, tobacco leaf oil is more, and quality is good, and judging parameter is preferable.As smoking and the attention rate of health problem improve, reducing tar and reducing harm is aobvious
It must become more and more important, it is also higher and higher to the quality requirements of tobacco leaf.In order to promote the quality of tobacco leaf, increases varietal resistance, improve leaf
Face secretion content is a very effective approach.
The study found that for tobacco, trichome density and glandular hairs type and the number of blade face secretion content are closely related.
Trichome density is bigger, and especially secreting type trichome density is bigger, and blade face secretion content is higher.And the wound of hypersecretion profile material
System, conventional breed breeding are extremely difficult to purpose, and what application was more at present is to carry out mutagenesis by the methods of physics, chemistry,
Objective trait is screened in mutant materials.Since the mutant materials frequency of mutation is low and the randomness of mutation is very strong, meet objective trait
Material it is considerably less, need to screen in large quantities of materials to obtain objective trait.The general screening side of hypersecretion profile material
Method is to carry out secretion to all processed single-strain plantings and one by one to extract identification, but the plantation of large batch of material and blade face point
Secretion extracting and developing, detection etc., it is time-consuming and laborious, cost is very high, workload is very big.Therefore, it finds a kind of simple and effective
Screening technique reduces cost to reduce workload, improves screening efficiency and is of great significance.
Summary of the invention
It formulates mainly for existing hypersecretion type tobacco-containing material using to the plantation of mutagenic progeny whole and to blade face one by one
Secretion extracts identification, and screening operation amount is big, at high cost, and the lower problem of efficiency, the purpose of the present invention is to provide one
The method that kind screens hypersecretion profile material from tobacco mutant body, can greatly reduce screening operation amount, and save the cost improves sieve
Select efficiency.
The present invention is realized by following technical proposals:
A method of screening hypersecretion profile material, main breeding process are as follows: seed treatment is (physico from tobacco mutant body library
Learn mutagenesis) preliminary screening → single plant screening and identification → repetitive identified (F2 generation identification) → ore grade indexes → production of → mutagenized populations
Test.
Wherein, comprising the following steps:
Step 1, seed treatment: the method for Applied Physics or chemistry handles with known characteristic tobacco seed, reaches mutagenesis
Purpose；
Step 2, the preliminary screening of mutagenized populations: by processed seed program request on the seedlings nursing plate equipped with suitable matrix, simple grain
Program request carries out floating seedlings under preference temperature；When growing to 5-7 piece leaf, the sight of glandular hairs morphology is carried out to the blade of seedling
It examines；The material that compared with wild type gland head is big and trichome density obviously increases is filtered out, while the single plant of clip is marked,
Or be grafted directly on a new drift disk, this process can eliminate 90% or more seedling；
Step 3, single plant screening and identification: single-strain planting is carried out in crop field to the target material that seedling screening goes out, while being planted wild
Type control.When the tobacco leaf maturity period, dyeing observation carried out to blade face glandular hairs, repetitive identified can secreting type glandular hairs density, and centering
Portion's leaf extracts blade face secretion according to standard method, carries out the detection of Related Component, according to testing result, retains blade face secretion
The higher material of content, single plant number, bagging are reserved seed for planting；
Step 4, repetitive identified (identification of F2 generation): F2 is planted for seed, field growth period eliminates grave illness strain and small and weak cigarette strain, choosing
Healthy and strong strain is selected, at the tobacco leaf maturity period, blade face secretion is carried out and extracts identification；And to its morphological feature, economical character, anti-
Characteristic of disease etc. carries out natural selection, and single plant system bagging is reserved seed for planting, and obtains the plant of more variation types as far as possible；
Step 5, ore grade indexes: the material obtained to F2 generation carries out variety test of lines, produces to the stabilization strain selected
Availability comparative evaluation determines target hypersecretion profile material.
Preferably, with any pair of cigarette in ethylmethane sulfonate (EMS), dithyl sulfate (DES), fast neutron in step 1
Grass seed is handled.
Preferably, glandular hairs morphological observation is carried out to the tobacco leaf of seedling in step 2 specifically: to its length to 5-7 piece
Ye Shi selects the blade of same size on seed plate, in the long blade of same position clip 0.5cm or so width, 2cm or so, protects
It holds fresh and alive, is unable to dehydration, is placed directly under super depth of field stereomicroscope and is observed.
Compared with the prior art, the invention has the following beneficial effects:
(1) it is provided by the invention from tobacco mutant body library screen hypersecretion profile material method, identification initial stage, not directly from
More complicated blade face secretion, which is started with, to be identified and is screened, but utilizes the phase of trichome density and type and blade face secretion
Guan Xing first carries out the screening of the density and type of intuitive blade face glandular hairs, then carries out the identification of blade face secretion on this basis
With the screening of hypersecretion profile material, by the screening of glandular hairs twice, secretion is identified twice, it is ensured that screens the Objective of offspring
Shape, the selection result are accurate and reliable；
(2) method provided by the invention that hypersecretion profile material is screened from tobacco mutant body library, has just eliminated 90% in Seedling Stage
Above mutant plants greatly reduce the plantation amount of crop field material, reduce the expenses such as land used, recruitment；
(3) method provided by the invention that hypersecretion profile material is screened from tobacco mutant body library, because of the reduction of planting material,
Reduce the quantity of maturity period secretion identification, the extraction of secretion and detection are time-consuming, the extraction of each sample, concentration,
Detection time in 3-5 hour, testing cost at 1000 yuan or so, the present invention largely reduce detection plant quantity (90% with
On), great amount of cost has been saved, screening time is shortened, has substantially increased screening efficiency.
Detailed description of the invention
Fig. 1 is wild type K326 glandular hairs morphologic observation photo.
Fig. 2 is T29 glandular hairs morphologic observation photo.
Fig. 3 is T12 glandular hairs morphologic observation photo.
Fig. 4 is T2 glandular hairs morphologic observation photo.
Specific embodiment
Further to illustrate technological means and its effect adopted by the present invention, below in conjunction with drawings and the specific embodiments
It is described in detail.
Embodiment 1
A method of screening hypersecretion profile material from tobacco mutant body library, comprising the following steps:
Step 1, seed treatment: the selection higher tobacco K326 seed of germination percentage uses dithyl sulfate according to debita spissitudo
(DES) it is handled, obtains mutagenesis seed.
Step 2, the preliminary screening of mutagenized populations: by processed seed program request on the seedlings nursing plate equipped with suitable matrix,
Selection retains 5000 plants of young shoot of energy normal growth after emergence, under preference temperature, carries out floating seedlings growth.It is long to 5- to seedling
When 7 leaves, drift disk is moved out control water, blade of the same size is selected on seed plate, in leaf central part clip 0.5cm or so width, 2cm
The long blade in left and right keeps fresh and alive, is unable to dehydration, is placed directly under super depth of field stereomicroscope, carries out glandular hairs morphological observation.
Compared with control wild type K326, gland head is significantly increased and material that trichome density obviously increases is taken pictures, while is right
The single plant of clip is marked.Biggish 12 plants of gland head are selected, increased 53 plants of density, number is T1 to T65.Fig. 1
For wild type K326 glandular hairs morphologic observation photo, Fig. 2 ~ 4 are respectively T29, T12, T2 glandular hairs morphologic observation photo.
Step 3, single plant screening and identification: the target material and wild type control go out to preliminary screening carries out strain kind in crop field
It plants, plants 66 parts altogether.Each strain plants 10 plants.When the maturity period, heavier, the weaker material of growing way, to 47 of falling ill is eliminated
The middle leaf of material, is dyed, under the microscope secondary observation, and extracts blade face secretion according to standard method, carries out leaf
Secrete analyte detection in face.According to the observation and testing result, retain higher 30 plants of material of blade face secretion content, single plant bagging stays
Kind.
Step 4, repetitive identified (identification of F2 generation): F2 is planted for seed according to conventional cultivation method, in the maturity period pair
Portion's leaf repeats the identification of blade face secretion, and carries out natural selection, single plant to its morphological feature, economical character, disease resistance etc.
Bagging is reserved seed for planting, and obtains the plant of more variation types as far as possible.By comparison, the material that 10 parts of secretion total amounts are more than 50% is obtained
Material, secretion content are shown in Table 1.The comprehensive agronomy character of wherein T12, T29, T31, T15, T23, this 5 materials are preferable.
1 F2 of table is for blade face secretion content table
Step 5, ore grade indexes: by obtained F3 for material, the preferable 5 strain T12, T29 of comprehensive agronomy character, T31, T15,
T23 carries out variety test of lines, mainly carries out the comprehensive comparative evaluations such as yield, the output value, disease resistance to the stabilization strain selected,
Screening obtains target strain T12, T29 that Comprehensive Traits are preferable, and blade face secretion dramatically increases.
From the result of embodiment 1 it is found that successfully having been filtered out from tobacco mutant body library using method of the invention more
Kind Comprehensive Traits are preferable, the hypersecretion profile material that blade face secretion dramatically increases, and this method is easy to operate, screening time is short, knot
Fruit is reliable, screening cost can be greatly reduced.
The above description is merely a specific embodiment, but scope of protection of the present invention is not limited thereto, ability
Other modifications or equivalent replacement that domain those of ordinary skill makes technical solution of the present invention, without departing from skill of the present invention
The spirit and scope of art scheme, are intended to be within the scope of the claims of the invention.
Claims (3)
1. a kind of method for screening hypersecretion profile material from tobacco mutant body library, which comprises the following steps:
Step 1, seed treatment: the method for Applied Physics or chemistry handles with known characteristic tobacco seed, reaches mutagenesis
Purpose；
Step 2, the preliminary screening of mutagenized populations: by processed seed program request on the seedlings nursing plate equipped with suitable matrix, simple grain
Program request carries out floating seedlings under preference temperature；When growing to 5-7 piece leaf, the sight of glandular hairs morphology is carried out to the blade of seedling
It examines；The material that compared with wild type gland head is big and trichome density obviously increases is filtered out, while the single plant of clip is marked,
Or be grafted directly on a new drift disk, this process can eliminate 90% or more seedling；
Step 3, single plant screening and identification: single-strain planting is carried out in crop field to the target material that seedling screening goes out, while being planted wild
Type control；When the tobacco leaf maturity period, dyeing observation carried out to blade face glandular hairs, repetitive identified can secreting type glandular hairs density, and centering
Portion's leaf extracts blade face secretion according to standard method, carries out the detection of Related Component, according to testing result, retains blade face secretion
The higher material of content, single plant number, bagging are reserved seed for planting；
Step 4, repetitive identified: F2 is planted for seed, field growth period eliminates grave illness strain and small and weak cigarette strain, selects healthy and strong strain
System carries out blade face secretion and extracts identification at the tobacco leaf maturity period；And its morphological feature, economical character, disease resistance etc. are carried out
Natural selection, single plant system bagging are reserved seed for planting, and obtain the plant of more variation types as far as possible；
Step 5, ore grade indexes: the material obtained to F2 generation carries out variety test of lines, produces to the stabilization strain selected
Availability comparative evaluation determines target hypersecretion profile material.
2. the method according to claim 1 for screening hypersecretion profile material from tobacco mutant body library, which is characterized in that step
It is handled in rapid 1 with any pair of tobacco seed in ethylmethane sulfonate (EMS), dithyl sulfate (DES), fast neutron.
3. the method according to claim 1 for screening hypersecretion profile material from tobacco mutant body library, which is characterized in that step
Glandular hairs morphological observation is carried out to the tobacco leaf of seedling in rapid 2 specifically: when its length to 5-7 piece leaf, select on seed plate
The blade of same size keeps fresh and alive, is unable to dehydration, is placed directly in super in the blade of same position clip 0.5cm or so width
It is observed under depth of field stereomicroscope.
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111109079A (en) * | 2020-01-21 | 2020-05-08 | 河南中烟工业有限责任公司 | Method for screening low-potassium-resistant tobacco variety mutant |
| CN111109078A (en) * | 2020-01-21 | 2020-05-08 | 河南中烟工业有限责任公司 | Method for screening high-aroma tobacco variety mutant |
| CN111448983A (en) * | 2020-01-21 | 2020-07-28 | 河南中烟工业有限责任公司 | Method for screening high-nicotine tobacco variety mutant |
-
2018
- 2018-11-19 CN CN201811374489.3A patent/CN109220786A/en not_active Withdrawn
Non-Patent Citations (1)
| Title |
|---|
| 李雪君等: "EMS 诱变不同香韵类型烟草突变体的初步筛选", 《河南农业科学》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111109079A (en) * | 2020-01-21 | 2020-05-08 | 河南中烟工业有限责任公司 | Method for screening low-potassium-resistant tobacco variety mutant |
| CN111109078A (en) * | 2020-01-21 | 2020-05-08 | 河南中烟工业有限责任公司 | Method for screening high-aroma tobacco variety mutant |
| CN111448983A (en) * | 2020-01-21 | 2020-07-28 | 河南中烟工业有限责任公司 | Method for screening high-nicotine tobacco variety mutant |
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