CN101049089A - Breeding method for filtering out resistance crop through adversity qualification - Google Patents
Breeding method for filtering out resistance crop through adversity qualification Download PDFInfo
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- CN101049089A CN101049089A CN 200710099574 CN200710099574A CN101049089A CN 101049089 A CN101049089 A CN 101049089A CN 200710099574 CN200710099574 CN 200710099574 CN 200710099574 A CN200710099574 A CN 200710099574A CN 101049089 A CN101049089 A CN 101049089A
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Abstract
A stress discriminating and screening method for culturing the stress-resistant crop (cotton) includes such steps as dividing the cotton specimen into two parts, culturing one in the non-stress condition, culturing another in the stress condition, comparing their relative characters with each other, and discriminating and screening the cotton variety with excellent characters.
Description
Technical field
The present invention relates to field of plant breeding, specifically identify by the crop adverse circumstance crop breeding offspring is selected and purifying, to quicken breeding process, improve efficiency of selection and accuracy, the inventive method is specially adapted to cotton breeding.
Background technology
The crop breeding of new variety lasts length, and breeding efficiency is low.The seed selection accuracy rate of antagonism kind is low, and the improved variety disease resistance is poor, and output is lower.
Cotton is most important in the world economic crops, also is the important source of China's foreign exchange earning.China just begins cotton breeding as far back as the forties in 20th century, but because breeding technique falls behind, mainly by the Mount Tai word cotton variety of line breeding transformation from U.S.'s introduction, makes slow progress at that time.Carry out crossbreeding after the fifties, the cotton field disease is light, basically according to output performance seed selection kind.Cotton wilt takes place to increase the weight of gradually after the seventies, begun the anti-blight breeding, proposition susceptible variety such as Northwest Agricultural University's height becomes are forever being identified the theory of selecting disease-resistant variety sickly, but breeding technique adopts the susceptible experimental field of nature to carry out disease resistance basically and identifies, because the pathogen deficiency, the field morbidity is inhomogeneous, and the time differences is big, breeding parent and filial generation material qualification result are inaccurate, and material is elected or superseded blindness is bigger.Verticillium wilt takes place gradually serious after the nineties, the particularly generation of defoliation verticillium wilt, generally to yield effect 20%-30%, press for and cultivate the resisting verticillium kind, but disease resistance identifies still and adopts and the similar method of anti-blight that breeding efficiency is low, and progeny material is selected inaccurate, the improved variety disease resistance is poor, and disease-resistant variety output is lower.After entering 21 century, the Cotton Production fast development, output increases sharply, the transgenic breeding technology is popularized rapidly, require variety yield, disease resistance, resistance to insects, drought-resistant, saline alkali and fiber quality to improve synchronously, therefore set up important content and an urgent demand that cotton resistance authenticate technology becomes the cotton breeding technological innovation.Transformation is disease-resistant, the molecular markers for identification technology of drought resisting and anti-saline and alkaline gene breeding is not appeared in the newspapers both at home and abroad, during domestic resistance to insects at transgenic cotton against pests is identified, the general authentication method that after the cotton final singling, adopts kanamycin to smear true leaf 1 time, not only operating efficiency and efficiency of selection are low, and bigger to the experimental field yield effect.
Summary of the invention
The objective of the invention is to provide a kind of breeding method of filtering out resistance crop through adversity qualification at above-mentioned deficiency, be used for to the crop breeding offspring select with purifying to quicken breeding process, improve efficiency of selection and accuracy rate, shorten breeding cycle, the inventive method is specially adapted to cotton breeding.
The inventive method is divided into two parts with specimen material to be identified, cultivates in non-adverse circumstance and adverse circumstance environment respectively, and contrast is set, and its resistance trait and productivity are investigated in the same management of cultivation simultaneously, filter out good kind.
Specifically, the present invention comes the evaluation and screening specimen material by production loss rate and/or volume increase percentage, and the computational methods of wherein said production loss rate are:
Described volume increase percentage comprises volume increase percentage under the non-adverse environmental factor and the volume increase percentage under the adverse environmental factor, and computational methods are under the non-adverse environmental factor:
Computational methods under the adverse environmental factor are:
Preferably selected specimen material judge index is volume increase percentage＞0, production loss rate≤20%.
The inventive method is specially adapted to the filial generation of transfer-gen plant genetic stability is screened, and does not see similar report at present both at home and abroad.
The present invention also provides and has identified and screened above-mentioned transfer-gen plant method, it is to adopt the NPTII genetic transformation plant that serves as a mark, identify and screen positive transformed plant by spraying the kanamycin aqueous solution, until the transfer-gen plant that obtains inheritance stability, the concentration of the described kanamycin aqueous solution is 2000～4000mgL
-1
Transgenic cotton against pests generally adopts NPTII both at home and abroad at present, it is the marker gene that generally adopts in the present crop breeding field, especially for cultivating transgenic cotton against pests, this gene pairs kanamycin produces the resistance reaction, the non-transgenic cotton is at the blade spraying kanamycin after 1 week, blade produces macula lutea, identifies in view of the above whether transgenic cotton has functional gene and express.In the evaluation to the NPTII marker gene, generally adopt kanamycin to smear blade both at home and abroad, the present invention adopts the kanamycin spray-on process of debita spissitudo to identify, identifies that efficient improves more than 50 times.Other all can adopt this technology as disease-resistant, drought resisting, anti-external source genetically modified crops such as saline and alkaline.Period is selected seedling stage and flowering and boll-setting period in evaluation, and identification traits is the transgenic line that the NPTII mark is arranged, comprise change pest-resistant, disease-resistant, drought resisting, anti-gene such as saline and alkaline all can adopt this technology.Authentication method: spray kanamycin by the field, the non-transgenic plant leaf produces macula lutea, and transfer-gen plant does not have this reaction.When crop grows 1 true leaf, use the atomizers spray kanamycin aqueous solution 1 time, flowering and boll-setting period repeats to spray 1 time.
According to the method described above genetically modified crops progeny population and offspring thereof are carried out continuous evaluation and screening, until the resistant plant that obtains genetic stability.Filter out the high material of value by output loss rate and volume increase percentage then.
Contrast of the present invention can be the strain with relevant antagonism, or the parent of breeding.
Transgenic pest-resistant cotton varieties such as middle cotton institute 30,31,32,41 and middle cotton institute 57 have successively been bred by the inventive method, breeding cycle shortens about 3 years than routine techniques, and the kind bollworm resisting of breeding is stablized the output height by force, wide adaptability, remarkable in economical benefits.
Embodiment
Following embodiment is used for further specifying of the present invention, but is not used for limiting the scope of the invention.
The cultivation of embodiment 1 transgene cotton and adverse circumstance evaluation and screening method
2003 with middle cotton institute 41 (state careful cotton 2002001, middle cotton seed industry science and technology limited Company) as the parent, with the cotton strain 957182 of routine (middle cotton seed industry science and technology limited Company) hybridization, the winter first familiar generation carried out south numerous added-generation in Sanya, Hainan in 2003, did not identify selection.2004 in Earthquake of Anyang station in Henan middle cotton institute experimental field plantation F2 generation, and field sowing is emerged the back cotton when growing 1 true leaf, and with the atomizers spray 2500ppm kanamycin aqueous solution 1 time, flowering and boll-setting period repeats to spray 1 time.Spray after 7～10 days for the 1st time, thoroughly pull out the non-transgenic positive plant of macula lutea, do not influence field final singling and other operation, can guarantee the density that the cotton field is enough.After spraying for the 2nd time, find that indivedual blades have the non-transgenic plant of macula lutea reaction, make mark with plastic ties or plastics board, can not influence experimental field output as assorted flower results harvest time, only selects unresponsive transfer-gen plant.Repeat said method its offspring is carried out the step sizing evaluation, eliminate bad combination and non-transgenic plant early, stable until other main character inheritance, till the offspring no longer separates.Obtain the transgenic line of 6 inheritance stabilities through screening, numbered respectively.
Inventor in 2005 carries out production loss comparison method (according to production loss rate and volume increase percentage) to 6 transgenic strains that obtain and identifies resistance to insects and value, test is established normal control cotton bollworm and is not is not prevented and treated two processing of cotton bollworm, and result of the test is as follows:
Table 1 production loss comparison method lint yield qualification result (kilogram/mu)
| Family name | Non-adverse circumstance | Adverse circumstance | The production loss rate | Rate of growth (non-adverse circumstance) | Rate of growth (adverse circumstance) |
| 501001 501002 501003 501004 501005 501006 | 98.5 86.4 77.5 99.2 85.3 69.4 | 87.5 53.2 68.5 90.2 81.4 58.7 | 11.16 38.42 11.61 9.07 4.57 15.41 | 11.04 -2.59 -12.62 11.83 -3.83 -21.75 | 8.56 -33.99 -15.01 11.91 0.99 -27.17 |
| Middle cotton institute 41 | 88.7 | 80.6 | 9.13 |
As can be seen from the above table, be numbered 501004 strain, its production loss rate is low, and increase yield significantly has good economic use value.
Embodiment 2
Process for screening and identifying according to embodiment 1, the inventor obtained transgenic pest-resistant cotton material R93-6, and (state examined cotton 980002 from 1998, middle cotton seed industry science and technology limited Company) since, being that centering is low carries out the kanamycin evaluation continuously for the breeding material bollworm resisting, annual after field emergence, with the atomizers spray 2500ppm kanamycin aqueous solution, react according to the anti-kanamycin of field plant after 1 week, before final singling, in time dial and remove the non-pest-resistant plant that produces macula lutea, 2 generation cotton bollworms (being generally June 20) spray once when taking place again, evaluation and screening technique be the first time together, and by production loss rate and volume increase percentage screening high yield strain is simultaneously.Utilizing this technology to carry out the generation continuously identifies, successively bred middle cotton institute 30,31,32,41 and transgenic pest-resistant cotton variety such as middle cotton institute 57 (therefrom cotton seed industry science and technology limited Company is bought, Earthquake of Anyang station in Henan), breeding cycle shortens about 3 years than routine techniques, the kind bollworm resisting of breeding is stablized by force, the output height, wide adaptability, popularizing area is big, remarkable in economical benefits, wherein pair valency transgenic cotton against pests middle cotton institute 41 is in Henan, Hebei, Shandong, Anhui, Shanxi, 2,300 ten thousand mu of Shaanxi Liu Sheng accumulative total popularizing areas, 3,000,000,000 yuan of newly-increased economic results in society obtained Chinese Academy of Agricultural Sciences's scientific and technological achievement first prize and Shaanxi Province's scientific and technological progress first prize in 2006.
Claims (8)
1, the breeding method of filtering out resistance crop through adversity qualification comprises the steps: specimen material to be identified is divided into two parts, cultivates in non-adverse circumstance and adverse circumstance environment respectively, and contrast is set, its resistance trait and productivity are investigated in the same management of cultivation simultaneously, filter out good kind.
2, the method for claim 1 is characterized in that, comes the evaluation and screening specimen material by production loss rate and/or volume increase percentage, and the computational methods of wherein said production loss rate are:
Described volume increase percentage comprises volume increase percentage under the non-adverse environmental factor and the volume increase percentage under the adverse environmental factor, and computational methods are under the non-adverse environmental factor:
Computational methods under the adverse environmental factor are:
3, method as claimed in claim 2 is characterized in that selecting volume increase percentage＞0, the specimen material of production loss rate≤20%.
As each described method of claim 1～3, it is characterized in that 4, described specimen material is the filial generation of genetic stability.
5, method as claimed in claim 4, it is characterized in that, the filial generation of described genetic stability is to obtain in the following way: adopt the NPTII genetic transformation plant that serves as a mark, screen positive transformed plant by spraying the kanamycin aqueous solution, until the transfer-gen plant that obtains inheritance stability.
6, method as claimed in claim 5 is characterized in that, the concentration of the described kanamycin aqueous solution is 2000～4000mgL
-1
As each described method of claim 1～3, it is characterized in that 7, described crop is a cotton.
8, as each described method of claim 1～3, it is characterized in that described resistance comprises resistance to insects, disease resistance, drought resistance, saline-alkali tolerance.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174570A (en) * | 2011-03-17 | 2011-09-07 | 西南大学 | Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same |
| CN102174571A (en) * | 2011-03-17 | 2011-09-07 | 西南大学 | Method for culturing anti-greensickness cotton by using artificially synthesized antimicrobial peptide gene |
| CN102613017A (en) * | 2012-03-04 | 2012-08-01 | 中国农业科学院棉花研究所 | Identification method for salt resistance of cotton blades |
| CN104429933A (en) * | 2014-09-22 | 2015-03-25 | 中国农业科学院棉花研究所 | Method for screening semi-wild cotton material with high salt and alkali resistance |
| CN105830758A (en) * | 2016-03-30 | 2016-08-10 | 中棉种业科技股份有限公司 | Method for identifying premature senescence resistance of cotton |
| CN116138161A (en) * | 2022-09-08 | 2023-05-23 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Crop drought-resistant breeding screening test method |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1099231C (en) * | 2000-11-03 | 2003-01-22 | 南京农业大学 | Breeding process for gene transfering cotton hybrid resisting bollworm |
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2007
- 2007-05-24 CN CN 200710099574 patent/CN101049089B/en not_active Expired - Fee Related
Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102174570A (en) * | 2011-03-17 | 2011-09-07 | 西南大学 | Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-greensickness cotton by using same |
| CN102174571A (en) * | 2011-03-17 | 2011-09-07 | 西南大学 | Method for culturing anti-greensickness cotton by using artificially synthesized antimicrobial peptide gene |
| CN102174571B (en) * | 2011-03-17 | 2012-09-05 | 西南大学 | Method for culturing anti-greensickness cotton by using artificially synthesized antimicrobial peptide gene |
| CN102174570B (en) * | 2011-03-17 | 2013-03-13 | 西南大学 | Plant expression vector for controlling artificially synthesized antimicrobial peptide gene by using specific vascular promoter and method for culturing anti-verticillium wilt cotton by using same |
| CN102613017A (en) * | 2012-03-04 | 2012-08-01 | 中国农业科学院棉花研究所 | Identification method for salt resistance of cotton blades |
| CN104429933A (en) * | 2014-09-22 | 2015-03-25 | 中国农业科学院棉花研究所 | Method for screening semi-wild cotton material with high salt and alkali resistance |
| CN105830758A (en) * | 2016-03-30 | 2016-08-10 | 中棉种业科技股份有限公司 | Method for identifying premature senescence resistance of cotton |
| CN116138161A (en) * | 2022-09-08 | 2023-05-23 | 广西壮族自治区亚热带作物研究所（广西亚热带农产品加工研究所） | Crop drought-resistant breeding screening test method |
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