CN105494079A - Method for acquiring cotton plants with high insect resistance - Google Patents
Method for acquiring cotton plants with high insect resistance Download PDFInfo
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- CN105494079A CN105494079A CN201510880361.4A CN201510880361A CN105494079A CN 105494079 A CN105494079 A CN 105494079A CN 201510880361 A CN201510880361 A CN 201510880361A CN 105494079 A CN105494079 A CN 105494079A
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- cotton
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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
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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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
Abstract
The invention discloses a method for acquiring cotton plants with high insect resistance. The method comprises steps as follows: a kanamycin sulfate solution with the concentration of 2,500-3,000 ppm is sprayed to leaf surfaces after first true leaves grow flatly at the early cotton emergence stage, and a single plant without yellow spots on the leaf surfaces of cotyledon and the true leaves is selected as a first processed plant after 6-8 days; when 6 true leaves grow on the first processing plant and the top first true leaf spreads flatly, the top third leaf and the top fourth leaf are inoculated with cotton bollworms, three 1-age larvae of the cotton bollworms are inoculated on each leaf, the leaf nibbling condition is observed after inoculation in 4-6 days, and cotton plants with high insect resistance, moderate resistance and low resistance are judged and screened according to the leaf nibbling degree and the death number of the larvae. According to the method, a cotton material with high insect resistance can be screened effectively, the insect resistance of the cotton is improved, the effective insect resistant performance of insect-resistant cotton is improved, induced tolerance of cotton with inefficient insect resistance on the cotton bollworms is avoided, the tolerance evolution process of the cotton bollworms on the insect-resistant cotton is reduced and eliminated, the usage quantity of related pesticides is reduced, and pollution is reduced.
Description
Technical field
The invention belongs to agricultural biological technical field, be specifically related to a kind of method obtaining high resistance to insects cotton plants.
Background technology
As everyone knows, the application of Insect Resistant Cotton has greatly promoted the development of Cotton in China industry, plays an important role to the development and stabilization of the industrial chain that Cotton in China is produced and it is correlated with.But due to composite factor impacts such as the characteristic of the Constantly allogamous of cotton and production managements, usually cause its resistance to insects lengthen along with cycle of plantation and weaken, cause unnecessary loss.
For above problem, also using many methods in prior art, spraying as adopted, the means such as to smear use kanamycin evaluation and screening Insect Resistant Cotton, reject that cotton plant of not anti-card, retain that plant of anti-card.But its in the process used because concentration standard is different, the difference on sprinkling opportunity, and the tolerance degree of cotton material itself, even artificial sprays the reasons such as custom, resistance can be obtained impure, namely there is false-positive plant, thus cause the accuracy deviation to some extent of result, affect the stable of cotton material resistance to insects.Connecing worm by biology carrys out evaluation and screening Insect Resistant Cotton in addition, if evaluation method for cotton bollworm resistance of Bt-transgenic cotton (CN201210339761) is exactly determine whether Insect Resistant Cotton by connecing worm by the dead degree of larva at present.Various method is only judge the presence or absence of resistance to insects above, such judgement no doubt can maintain the uniformity of Insect Resistant Cotton, but owing to not judging the power of resistance to insects further, the later use to material can be caused, so the validity of its production application management etc. and accuracy deviation to some extent.Therefore how obtaining high resistance to insects cotton plants and effectively being transformed and utilize is the problem being worth research and exploring.
Summary of the invention
The object of the present invention is to provide a kind of method that effectively can improve cotton breeding parent material resistance to insects, thus obtain the cotton material of high resistance to insects.
For reaching above object, the invention provides a kind of method obtaining high resistance to insects cotton plants, the method comprises the following steps:
(1) emerge the initial stage in cotton, after the 1st true leaf growth of cotton is open and flat, with the kanamycin sulfate spray solution blade face that concentration is 2500-3000ppm, the blade choosing individual plant cotyledon and true leaf after 6-8 days does not occur that the plant of yellow spotting is the first process plant;
(2) treat that the first process plant strain growth is to there being 6 true leaves, and fall 1 true leaf open and flat after, get down 3 and fall 4 leaves inoculation cotton bollworms, every sheet blade is inoculated cotton bollworm 1 instar larvae 3, inoculate and observe blade after 4-6 days and nibble situation, according to blade degree of nibbling and dead larvae head number judge to filter out high resistance, in anti-and anti-type cotton plant.
Wherein, after entering the flower bud florescence, can by the plant of high resistance and in the flower of anti-plant all do selfing, selfing bell is received according to the every strain list of numbering, selfing bell is carried out Nan Fan by the end of the year, and the selfing bell plantation of every strain is embarked on journey, and often row continues to do selfing, individual plant is divided to collect and number, Second Year plants 2 row according to high resistance selfing bell, in anti-selfing bell plant 1 row, carry out 2:1:2:1 ratio line number intersect plant, during results, mixed receipts, carry out field planting by mixed sowing on the 3rd year and just can utilize further.
Wherein, relative to every mu of cotton in step (1), the consumption of kanamycin sulfate solution is 8-9 liter.
Wherein, in step (2), blade is placed on respectively and specially connects in worm container, preferably by described fall 3 and fall 4 leaves and be positioned in the bulge of high 3-5cm, diameter 4-6cm and inoculate cotton bollworm.
Wherein, judge in step (2) screening high resistance, in standard that is anti-and anti-type cotton plant as shown in table 1:
Table 1
One aspect of the present invention additionally provides the application of described method in cotton breeding.
Another aspect of the present invention additionally provides the application of described method in cotton bollworm control.
Process provides a kind of method of screening and raising bt-cotton (mainly bollworm resisting), use the method effectively can reject not pest-resistant and that resistance to insects is more weak cotton material, the cotton material that screening resistance to insects is strong, thus improve the resistance to insects of cotton, promote the pest-resistant effective performance of Insect Resistant Cotton, avoid the pest-resistant inducing tolerance to cotton bollworm of poor efficiency, reduce and stop the tolerance evolution process of cotton bollworm to Insect Resistant Cotton, reduce the usage amount of relevant agricultural chemicals simultaneously, decreasing pollution, has positive production and social effect.
Embodiment
Below in conjunction with embodiment, the present invention is described in detail.
Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.If do not specialize, the conventional means that in embodiment, technological means used is well known to those skilled in the art, is raw materials usedly commercial goods.
Embodiment 1
2009 to 5017 (CCRI 41 × CCRIs 17) in material, in 6416 (CCRI 52 × CCRIs 43) when carrying out Continuous Observation, find that field blade has to nibble, cotton buds and bolls also had and nibbled loss the cotton buds and bolls phase, within 2010, utilize convenient technical process, receive after mycin Screening and Identification by concentration 3000ppm card, to do not react plant carry out field toxalbumin content detection time, discovery effect is undesirable, testing result is lower than pest-resistant contrast, due to two resistance to sick high-qualitys of material, value is higher, need to screen raising further, 2010 the end of the year results do not react plant and carry out southern breeding and plant, what plantation in 2011 was bred does not react plant, after the 1st true leaf growth of cotton is open and flat, kanamycin screening is carried out with the spray kanamycin sulfate solution that concentration is 3000ppm, fountain height is 8-9 liter/mu, the blade choosing individual plant cotyledon and true leaf after 7 days does not occur that the plant of yellow spotting is the first process plant, treat that the first process plant strain growth is to there being 6 true leaves, and fall 1 true leaf open and flat after, to fall 3 and fall 4 leaves be positioned over high 4cm, carry out in the bulge of diameter 5cm connecing worm screening, every sheet leaf connects 31 instar larvaes, after 4 days, in conjunction with the situation of pest-resistant and not pest-resistant control sample, (pest-resistant control sample blade has and nibbles to connect worm, 3 larvas are all dead, not pest-resistant control sample blade is nibbled area and is larger than over half, 2 above larvae alive of larva) just deployable investigation records, result classification (see table 1) is carried out according to record table, the result of combining classification is to high resistance, in anti-plant carry out selfing, each plant plantation in the end of the year 2011 is embarked on journey, within 2012, plant 2 row according to high resistance selfing bell, in anti-selfing bell plant the principle of 1 row, carry out 2:1:2:1 ratio line number intersection to plant, mixed receipts during results, mixed sowing was carried out field planting in 2013.The condition that manage same with pest-resistant contrast, estimates in field, and plant organ significantly reduces by worm harm, and field toxalbumin content detection is carried out in sampling simultaneously, and expression is significantly improved than contrast, have received good actual effect.(seeing the following form 2)
Table 2
Embodiment 2
Cotton 0913 [cotton No. 11 of (CCRI 35 × CCRI 69) × Henan] in the cotton hybrid generation kind of assembly in 2009, field multiple spot in 2010 is surveyed in product process, show disease-resistant stable yields, average effect of increasing production is obvious, but it is undesirable in the part test pilot effect that increases production and improves productivity, react through overtesting underwriter and find that main cause is the more weak instability of resistance to insects in conjunction with its variable rate technology, this material is a first-filial generation kind, its male parent is non-Insect Resistant Cotton, its anti insect gene derives from its female parent, therefore the resistance to insects improving its female parent just directly can effectively improve its resistance to insects.
First conventional rejuvenation pure keeping work is carried out to its male parent, secondly emphatically according to this method, screening is carried out to its maternal resistance to insects at the maternal merit of maintenance simultaneously and improve, be specially:
Emerging the initial stage first in cotton, after the 1st true leaf growth of cotton is open and flat, kanamycin qualification is carried out to test material, use the pure pulvis of kanamycin sulfate, the concentration 3000ppm (every 10000ml water adapted 30 grams of pulvis) used, after fully stirring evenly, artificial spray, fountain height is 8-9 liter/mu, after 7 days, with reference to the pest-resistant and not pest-resistant response situation contrasted, (pest-resistant contrast 100% is reactionless in dispenser, not pest-resistant contrast 100% is reacted) launch field investigation, pull out plant blade occurring yellow spotting, for blade not occurring the plant of yellow spotting retains and is numbered process of listing, numbering plant has grown to 6 true leaves, and fall 1 true leaf open and flat after, it is sampled, each cotton plant take down 3 and fall 4 leaves, to fall 3 and fall 4 leaves be positioned over high 4cm, carry out in the bulge of diameter 5cm connecing worm screening, cotton bollworm 1 instar larvae 3 is connected at every sheet leaf, by the time observation is started after connecing the 5th day of worm, (pest-resistant control sample blade has and nibbles the concrete situation with reference to pest-resistant and not pest-resistant control sample, 3 larvas are all dead, not pest-resistant control sample blade is nibbled area and is larger than over half, 2 above larvae alive of larva) just deployable investigation, specific standards is with reference to table 1.After existing cotton plant is classified, retain high resistance and in anti-type cotton plant and number, pull out simultaneously resistance to and sense type plant.
Enter the flower bud florescence, by high resistance and in anti-plant flower all do selfing, selfing bell is received according to the every strain list of numbering, the end of the year selfing bell will be planted and carry out Nan Fan in Hainan, the selfing bell plantation of every strain is embarked on journey, often row continues to do selfing, individual plant is divided to collect and number, Second Year plants 2 row according to high resistance selfing bell, in anti-selfing bell plant 1 row, carry out 2:1:2:1 ratio line number intersection to plant, mixed receipts during results, mixed sowing was carried out mixed again when field planting is gathered in the crops receipts in 3rd year, utilize mixed sowing as female parent and carry out the production of hybrid seeds, its first-filial generation is in same pilot, cotton bollworm larvae corrected mortality is higher, be better than contrast, and its average Bt insecticidal proteins expression (ng/g fresh weight) is also higher than contrast, show good resistance to insects, its yield level there has also been good raising simultaneously.(seeing the following form 3).
Table 3
Although above the present invention is described in detail with a general description of the specific embodiments, on basis of the present invention, can make some modifications or improvements it, this will be apparent to those skilled in the art.Therefore, these modifications or improvements without departing from theon the basis of the spirit of the present invention, all belong to the scope of protection of present invention.
Claims (6)
1. obtain a method for high resistance to insects cotton plants, it is characterized in that, the method comprises the following steps:
(1) emerge the initial stage in cotton, after the 1st true leaf growth of cotton is open and flat, with the kanamycin sulfate spray solution blade face that concentration is 2500-3000ppm, the blade choosing individual plant cotyledon and true leaf after 6-8 days does not occur that the plant of yellow spotting is the first process plant;
(2) treat that the first process plant strain growth is to there being 6 true leaves, and fall 1 true leaf open and flat after, get down 3 and fall 4 leaves inoculation cotton bollworms, every sheet blade is inoculated cotton bollworm 1 instar larvae 3, inoculate and observe blade after 4-6 days and nibble situation, according to blade degree of nibbling and dead larvae head number judge to filter out high resistance, in anti-and anti-type cotton plant.
2. method according to claim 1, is characterized in that, in step (1), relative to every mu of cotton, the consumption of kanamycin sulfate solution is 8-9 liter.
3. method according to claim 1 and 2, is characterized in that, in step (2), by described fall 3 and fall 4 leaves and be positioned in the bulge of high 3-5cm, diameter 4-6cm and inoculate cotton bollworm.
4. method according to claim 3, is characterized in that, judge in step (2) screening high resistance, in standard that is anti-and anti-type cotton plant as shown in table 1:
Table 1
5. the application of the method in claim 1-4 described in any one in cotton breeding.
6. the application of the method in claim 1-4 described in any one in cotton bollworm control.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510880361.4A CN105494079A (en) | 2015-12-02 | 2015-12-02 | Method for acquiring cotton plants with high insect resistance |
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| CN201510880361.4A CN105494079A (en) | 2015-12-02 | 2015-12-02 | Method for acquiring cotton plants with high insect resistance |
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1373992A (en) * | 2002-04-28 | 2002-10-16 | 南京农业大学 | Breeding process of transgenic bollworm resisting cotton hybrid |
| US20110154523A1 (en) * | 2009-12-17 | 2011-06-23 | Pioneer Hi-Bred International, Inc. | Maize event DP-004114-3 and methods for detection thereof |
| CN102499060A (en) * | 2011-11-01 | 2012-06-20 | 江苏省农业科学院 | Transgenic helicoverpa armigera-resistant and phenol-free cotton breeding method |
-
2015
- 2015-12-02 CN CN201510880361.4A patent/CN105494079A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1373992A (en) * | 2002-04-28 | 2002-10-16 | 南京农业大学 | Breeding process of transgenic bollworm resisting cotton hybrid |
| US20110154523A1 (en) * | 2009-12-17 | 2011-06-23 | Pioneer Hi-Bred International, Inc. | Maize event DP-004114-3 and methods for detection thereof |
| CN102499060A (en) * | 2011-11-01 | 2012-06-20 | 江苏省农业科学院 | Transgenic helicoverpa armigera-resistant and phenol-free cotton breeding method |
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