CN103460923A - Method for identifying aphid resistance of wild cotton or semi-wild cotton - Google Patents
Method for identifying aphid resistance of wild cotton or semi-wild cotton Download PDFInfo
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Abstract
The invention relates to the field of identification of aphid resistance of cotton, in particular to a method for identifying aphid resistance of wild cotton or semi-wild cotton. The method comprises the following steps of: (1) selecting tender leaves at the tops of to-be-identified wild cotton or semi-wild cotton materials; (2) placing the tender leaves at the tops in a mesh enclosure; (3) carrying out manual aphid inoculating, inoculating 30 to 40 aphids for each leaf and sealing up a mesh bag mouth; (4) surveying the numbers of aphids of a checked variety and the numbers of aphids of a contrast variety; (5) calculating breeding magnification of aphids. The method disclosed by the invention eliminates interference of factors such as other insect pests, natural enemies and the like and has a stable and reliable identification result and strong practicality and operability.
Description
Technical field
The present invention relates to a kind of Resistance Strain of Cotton aphid and identify field, specifically, relate to the cotton aphis resistance authentication method of a kind of wild cotton or semi-wild.
Background technology
Although transgenic cotton against pests has effectively been checked the harm of cotton bollworm, cultivation contains from the natural Insect Resistant Cotton of anti insect gene in Gossypium still significant.In recent years, some researchers also are being devoted to utilize the landwards research of cotton transformation desirable genes of wild cotton, but have no, obtain germplasm valuable and that proterties is stable.
China cotton from Asian cotton, Bi Keshi, plucked instrument Bai Shi is cotton, cotton, abnormal cotton, the G.sturtianum of Si Tuokeshi, Somalia is cotton, Mexico's semi-wild is cotton, transformation bollworm resisting spy the wild species such as Bao Moershi is cotton, broad-leaved is cotton, thick and disorderly cotton, from Asian cotton, extremely cotton, wild species such as Si Tuokeshi is cotton, thick and disorderly cotton transformation anti-cotten aphid characteristic, be bred as totally 97 parts of pest-resistant external source blood relationship germplasms, the high resistance level that reached that the resistance to insects of this class germ plasm resource has reached anti-level, had.They are the good anti insect gene storehouses of cultivating nature Insect Resistant Cotton kind, also can be used for improveing transgenic cotton against pests.In sum, the Cotton in China distant hybridization is carried out the germ plasm resource creation and has been entered the international rank of advanced units, has reached gradually the practical stage that research of fruit germplasm resource utilizes.The height that distant hybridization produces forms the full content of the upland cotton germplasm with wild cotton foreign gene for germplasm line, distinguished germ plasm and some kinds or strain, no matter from germplasm quantity, or germplasm qualitatively, they all will produce far-reaching influence to cotton breeding from now on.
Present stage mainly relies on chemical insecticide to the control of cotton field cotten aphid, and along with a large amount of uses of insecticide certainly will cause cotten aphid to produce resistance to insecticide, finding new cotten aphid, to control approach very necessary.Current scientist does not also excavate and is cloned into the gene of effective control cotten aphid, and the cotton variety of cultivating the anti-cotten aphid of transgenosis also needs the time of growing.Pass through long-term domestication and directive breeding simultaneously, make the genetic diversity of cotton culture kind now more and more narrower.Yet, the edge kind far away such as other cultivated speciess outside the upland cotton cultivated species, L. Races, wild species, due to the environment of its distributed areas and long-term natural selection, make them there is abundant genetic diversity, contain the many excellent genes that can be applied by cotton breeding, as drought resisting, Resistant, cold-resistant, seed have potential characteristic that body of gland, male sterile and fiber finer are strong etc. without the body of gland plant.And natural Insect Resistant Cotton resistance to insects is from Gossypium, the strong target insect of insect-resistance is difficult for producing resistance to insects, and can not have a negative impact to environment and bio-safety.
After the transgenic cotton against pests establishing in large scale, the status of cotton pest develops, and the minor pests such as cotten aphid rise to primary pest, and it is caused harm and is day by day serious trend.Because former cotten aphid is not primary pest, and most of research resistance authentication method carries out round cultivar, attribute in the resistance of wild cotton or the cotton this ecosystem of semi-wild utilized less, or even blank.The cultivation cotton variety is therophytes, and germplasm resource is very abundant, can be as required any time any place as field, solarium's plantation after, take blade hazard level and aphid evil decline rate as identification of indicator be fully passable, but wild cotton and semi-wild cotton are the germplasm resource for cotton of life throughout the year or perennial preciousness, can not have a large amount of experiment samples to be used for carrying out the aphid resistance evaluation; Wild cotton and semi-wild cotton be all by potted plant growth near Zhong Huo laboratory, greenhouse, be not suitable for aphid naturally-occurring, can't carry out the whole strain of nature and identify; Even simultaneously because its preciousness has aphid, occur also can not wait until that blade extremely all crispaturaed and just can carry out the investigation of index by aphid damage; Under nature, China's wild cotton and semi-wild Gossypium kind are distributed in the Yunnan-Guizhou Plateau and cotton region, south China, these cotton regions are not the Cotton in China producing regions, and aphid generating capacity itself is just low, sample and preciousness thereof, understand its aphis resistance, we need be in the situation that do not destroy germplasm resource and identified.
The present invention studies discovery, adopts conventional method to identify the aphid resistance of cultivation cotton variety, and the aphid of take refers to that decline rate is as criterion, and result shows in 216 parts of materials, in 5 parts of anti-materials, 211 parts of resistant materials, without other resistance ranks.Therefore by result, can be found out, the aphid resistance authentication method that conventional cultivation grows cotton is not suitable for the aphid resistance authentication method of wild cotton and semi-wild cotton, the aspects such as wild cotton is extremely precious in addition, growing environment is special, take single leaf authentication method and aphid, to increase multiplying power more scientific as judging index.
Therefore be devoted to utilize the anti-cotten aphid kind material of screening in wild cotton or semi-wild cotton, and, by molecule labelling method, new material is created in polymerization, offered breeding man and use, become extremely urgent work.Therefore, be badly in need of the authentication method that research is applicable to wild cotton or the cotton anti-cotten aphid of semi-wild.
Summary of the invention
The purpose of this invention is to provide the cotton aphis resistance authentication method of a kind of wild cotton or semi-wild.
Therefore, peculiar characteristics according to wild cotton and semi-wild cotton self, the present invention has taken the lead in studying wild cotton or the semi-wild cotton control action to cotten aphid, and, by relatively field plot trial, solarium's test test, has researched and proposed the cotton aphis resistance authentication method of wild cotton or semi-wild.
According to wild cotton of the present invention or the cotton aphis resistance authentication method of semi-wild, comprise the following steps:
(1) select the top tender leaf of identified wild cotton or semi-wild cotton material; If the blade of selecting is improper, can affects the growth and breeding of aphid, thereby cause the evaluation error;
(2) the top tender leaf is placed in the guard of suitable size, stops the intrusion of insect and natural enemy; The aperture of mesh bag needs 100 orders, and aperture is too little, affects the blade normal growth, thereby causes that Leaf Physiology changes, and causes that aphid takes food variation, causes the evaluation error; Aperture is too large, and aphid can escape;
(3) manually connect aphid, every leaf is connected into aphid 30-40 head, seals the mesh bag mouth, prevents the aphid escape; It is crucial connecing the borer population amount, unsuitable too many, and too many breeding is too many, and survey error is large; Very little, breeding amount is inadequate in the aphid access;
(4) connect after aphid the 6th day, kind and check variety Aphed population are examined in investigation;
(5) calculate aphid breeding multiplying power, Aphed population on the analytic statistics different materials, and calculate aphid breeding multiplying power, aphid breeding multiplying power is the mean value of Aphed population on 10 strain cottons; Computing formula is as formula (1):
In formula (1):
A is the multiplying power that aphid breeds on wild cotton or semi-wild cotton;
X is the quantity of aphid while investigating on wild cotton or semi-wild cotton, and unit is " head ".
A is the inoculation Aphed population, and unit is " head ".
According to the specific embodiment of the present invention, the cotton aphis resistance authentication method of described wild cotton or semi-wild comprises:
(1) the test sheet leaf carries out, and selects respectively the top tender leaf of each material, and each single leaf is designated as once and repeats, and repeats for totally 10 times;
(2) will identify that this tender leaf of cotton is placed on the guard interior (100 order) of suitable size, stops the intrusion of insect and natural enemy;
(3) manually connect aphid, every leaf is connected into aphid 30-40 head, seals the mesh bag mouth, prevents the aphid escape;
(4) connect after aphid the 6th day, kind and check variety Aphed population are examined in investigation;
(5) calculate aphid breeding multiplying power;
(6) Aphed population on the analytic statistics different materials, and calculate aphid breeding multiplying power; Aphid breeding multiplying power is the mean value of Aphed population on 10 strain cottons; Computing formula is as formula (1):
In formula (1):
A is the multiplying power that aphid breeds on wild cotton or semi-wild cotton;
X is the quantity of aphid while investigating on wild cotton or semi-wild cotton, and unit is " head ".
A is the inoculation Aphed population, and unit is " head ".
(7) differentiate the resistance rank of identification of species according to aphid breeding multiplying power on different materials.
The cotton aphis resistance sex determination of table 1 wild cotton or semi-wild standard
| Resistance level | Aphid breeding multiplying power |
| High resistance | <1 |
| Anti- | 1～2 |
| In anti- | 2～4 |
| Low anti- | 4～5 |
| Sense | >5 |
Method of the present invention has been got rid of the interference of the factors such as other insects and natural enemy, and qualification result is stable, reliable, and practicality and workable is saved exotic material simultaneously.
Embodiment
Embodiment 1:
1, experimental technique:
(1) the test sheet leaf carries out, and selects respectively the top tender leaf of each material, and each single leaf is designated as once and repeats, and repeats for totally 10 times;
(2) will identify that this tender leaf of cotton is placed on the guard interior (100 order) of suitable size, stops the intrusion of insect and natural enemy;
(3) manually connect aphid, every leaf is connected into aphid 30-40 head, seals the mesh bag mouth, prevents the aphid escape;
(4) connect after aphid the 6th day, kind and check variety Aphed population are examined in investigation;
(5) calculate aphid breeding multiplying power.
(6) Aphed population on the analytic statistics different materials, and calculate aphid breeding multiplying power; Aphid breeding multiplying power is the mean value of Aphed population on 10 strain cottons; Computing formula is as formula (1):
In formula (1):
A is the multiplying power that aphid breeds on wild cotton or semi-wild cotton;
X is the quantity of aphid while investigating on wild cotton or semi-wild cotton, and unit is " head ".
30 is the inoculation Aphed population, and unit is " head ".
(7) the resistance rank of the differentiation of the breeding multiplying power on wild cotton or semi-wild cotton identification of species according to aphid.
2, experimental result:
From experimental result, single leaf evaluation and screening material amounts to 216 parts, and the data that investigation is gathered adopt DPS7.05 software LSD method to carry out multiple comparison analyse.The difference analysis result is as table 2, and result shows, the cotton aphis resistance criterion of wild cotton or semi-wild is judged to be with aphid breeding multiplying power:
High resistance: be less than 1; Anti-: 1～2; In anti-: 2～4; Low anti-: the 4-5 head; Sense: be greater than 5.
In 216 parts of materials of resistance standard determination, 1 part of high resistance material, 40 parts of resistant materials, in 99 parts of anti-materials, 32 parts of low anti-materials, 44 parts of abilities.Wherein CK is perceptual anti-aphid contrast cotton Africa E40, and the breeding multiplying power is 11.33, and the blade hazard level is 3.17, and result is basically identical, so this authentication method is accurate, feasible.
The representative qualification result of table 2 part
| Cotton material | The breeding multiplying power | Resistance is judged |
| In 2101 | 0.8800 | High resistance |
| Hubei Province cotton 23 | 1.0667 | Anti- |
| Shan cotton 2365 | 1.1233 | Anti- |
| Africa E40(CK) | 11.2333 | Sense |
Embodiment 2:
1, evaluation mode:
Utilize nature worm source, field, killed classification by inquiry, finally compare with check variety with damage index, the resistance level of evaluation different materials to seedling aphid and volt aphid.
2, field design:
The cotton sick lighter field that occurs of choosing, the community plantation, do not establish repetition.Every community 6 row, the long 8m of row, line-spacing 0.7-0.8m, spacing in the rows 26cm, 180Zhu， community area 33.6m altogether
2left and right.
Contrast is african cotton E-40.In local proper seeding, cotton seed is not made any anti-aphid chemicals treatment.Other cultivation management, with general land for growing field crops, does not increase any special measure.
3, investigation method:
Control time: betide cotton seedling tri-leaf period, five leaf phases (occurrence in peak period) and seven leaf phases the seedling aphid and carry out three investigation; In the volt aphid phase, carry out twice investigation.
Sampling method: middle four lines is got in every community, look at random 50 strains, twice investigation of seedling aphid phase all be take in the leaf of 5, cotton plant top and is injured the heaviest blade as the standard leaf be injured the 3rd this investigation of the heaviest blade, seedling aphid phase and twice investigation of volt aphid phase of complete stool, according to killed grade scale, adds up strain numbers at different levels.
After seedling aphid investigation for the second time, immediately with aphid of the pesticide controls such as nuvacron.
4, grade scale and calculating:
Aphid evil grade scale:
| Anti-level | Grade scale |
| Anti- | Aphid refers to that decline rate is greater than 25% |
| In anti- | Aphid refers to that decline rate is less than or equal to 25%, is greater than 10% |
| Middle sense | Aphid refers to that decline rate is less than or equal to 10%, is greater than-60% |
| Sense | Aphid refers to that decline rate is less than or equal to-60%, is greater than-80% |
| With contrast | Aphid refers to that decline rate is less than or equal to-80% |
5, experimental result:
From experimental result, adopt the aphid resistance of conventional cultivation cotton variety to identify that material amounts to 216 parts, the aphid of take refers to that decline rate is as criterion, result shows in 216 parts of materials, in 5 parts of anti-materials, 211 parts of resistant materials, without other resistance ranks.Therefore by result, can be found out, the aphid resistance authentication method that conventional cultivation grows cotton is not suitable for the aphid resistance authentication method of wild cotton and semi-wild cotton, the aspects such as wild cotton is extremely precious in addition, growing environment is special, take single leaf authentication method and aphid, to increase multiplying power more scientific as judging index.
The qualification result that table 3 part utilizes conventional method to obtain
| Cotton material | Aphid damage index (%) | Aphid refers to decline rate (%) | Resistance is judged |
| Inferior No. 1 of stone system | 65.63 | 17.1 | Anti- |
| HG-E-12 | 58.33 | 26.32 | Anti- |
| BRS-10 | 53.13 | 32.89 | Anti- |
| Acalasj-3 | 72.92 | 7.89 | Anti- |
Claims (3)
1. the cotton aphis resistance authentication method of a wild cotton or semi-wild, is characterized in that, said method comprising the steps of:
(1) select the top tender leaf of identified wild cotton or semi-wild cotton material;
(2) the top tender leaf is placed in the guard of suitable size, stops the intrusion of insect and natural enemy;
(3) manually connect aphid, every leaf is connected into aphid 30-40 head, seals the mesh bag mouth, prevents the aphid escape;
(4) connect after aphid the 6th day, kind and check variety Aphed population are examined in investigation;
(5) calculate aphid breeding multiplying power, Aphed population on the analytic statistics different materials, and calculate aphid breeding multiplying power, aphid breeding multiplying power is the mean value of Aphed population on 10 strain cottons; Computing formula is as formula (1):
In formula (1):
A is the multiplying power that aphid breeds on wild cotton or semi-wild cotton;
X is the quantity of aphid while investigating on wild cotton or semi-wild cotton, and unit is " head ".
A is the inoculation Aphed population, and unit is " head ".
2. the cotton aphis resistance authentication method of wild cotton according to claim 1 or semi-wild, is characterized in that, in step (1), each single leaf is designated as once and repeats, and repeats for totally 10 times.
3. the cotton aphis resistance authentication method of wild cotton according to claim 1 or semi-wild, is characterized in that, in step (2), the top tender leaf is placed in the guard of 100 order sizes.
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| CN103704000A (en) * | 2013-12-26 | 2014-04-09 | 董金锋 | Corn high-yield cultivation method |
| CN104012398A (en) * | 2014-06-13 | 2014-09-03 | 中国农业科学院棉花研究所 | Method for screening high-glyphosate-resistant semi-wild cotton material |
| CN104719013A (en) * | 2015-03-25 | 2015-06-24 | 中国农业科学院棉花研究所 | Wild cotton or semi-wild cotton mirid resistance identification method |
| CN111768044A (en) * | 2020-07-01 | 2020-10-13 | 石河子大学 | Method and system for monitoring cotton aphid number in seedling stage of cotton |
| CN114532175A (en) * | 2022-02-18 | 2022-05-27 | 新疆农垦科学院 | Method for identifying physiological aphid resistance based on cotton leaf downy characteristics |
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| CN101520447A (en) * | 2009-04-08 | 2009-09-02 | 南京农业大学 | Method for identifying aphis resistance of chrysanthemum by artificial inoculation |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103704000A (en) * | 2013-12-26 | 2014-04-09 | 董金锋 | Corn high-yield cultivation method |
| CN104012398A (en) * | 2014-06-13 | 2014-09-03 | 中国农业科学院棉花研究所 | Method for screening high-glyphosate-resistant semi-wild cotton material |
| CN104012398B (en) * | 2014-06-13 | 2016-05-04 | 中国农业科学院棉花研究所 | The method of screening high-resistance glyphosate semi-wild cotton material |
| CN104719013A (en) * | 2015-03-25 | 2015-06-24 | 中国农业科学院棉花研究所 | Wild cotton or semi-wild cotton mirid resistance identification method |
| CN111768044A (en) * | 2020-07-01 | 2020-10-13 | 石河子大学 | Method and system for monitoring cotton aphid number in seedling stage of cotton |
| CN111768044B (en) * | 2020-07-01 | 2022-07-08 | 石河子大学 | Method and system for monitoring cotton aphid number in seedling stage of cotton |
| CN114532175A (en) * | 2022-02-18 | 2022-05-27 | 新疆农垦科学院 | Method for identifying physiological aphid resistance based on cotton leaf downy characteristics |
| CN114532175B (en) * | 2022-02-18 | 2023-10-27 | 新疆农垦科学院 | Identification method for physiological aphid resistance based on fuzz characteristics of cotton leaves |
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Application publication date: 20131225 |