CN103329660A - Purity detection method for double-optimal 888 salt seeds of bilinear hybrid rice - Google Patents
Purity detection method for double-optimal 888 salt seeds of bilinear hybrid rice Download PDFInfo
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Abstract
The invention discloses a purity detection method for double-optimal 888 salt seeds of bilinear hybrid rice. By the method, according to a theory that after years of research, the salt tolerance of the double-optimal 888 salt seeds of the bilinear hybrid rice and the salt tolerance of the female parent 582S salt seeds of the bilinear hybrid rice are obviously different; under a certain salt (Nacl) concentration stress condition, the double-optimal 888 salt seeds and the 582S salt seeds (contrast) are cultured in a laboratory to obtain young seedlings; after the seedlings appear until 3-4 leaf periods, and if leaf tips are curled, leaves are withered, and the leaves slowly grow, the seeds are intolerant against the salt, and the seeds are the double-optimal 888 salt seeds; under the salt (Nacl) stress condition, if the leaves normally grow, the seeds are tolerant against the salt, and the seeds are solid seeds obtained by selfing of the female parent 582S salt seeds and the double-optimal 888 salt seeds in the last season; and therefore, the purity of the double-optimal 888 salt seeds can be calculated. By the method, the purity of the double-optimal 888 salt seeds of the bilinear hybrid rice can be quickly detected and the cost and the time of performing field planting on hybrid rice seeds in Hainan are saved.
Description
Technical field
The invention belongs to seed quality detection technique field, relate to the method for hybrid rice seeds purity detecting, a kind of double-linear hybrid rice salt two excellent 888 seed purity detection methods of more specifically saying so.
Background technology
Paddy rice is the first cereal crops of China, and China is maximum in the world hybrid rice seeds producing country and country of consumption, and producing highly purified hybrid rice seed all is the basis of giving full play to the hybrid vigour yield potential.In the recent period, the research and extension of double-linear hybrid rice has been made positive contribution for China's Rice Production, because the female parent of double-linear hybrid rice is photo-thermo-sensitive genic male sterile line, photo-thermo-sensitive genic male sterile line enters the influence that is subjected to environmental conditions such as illumination, temperature early stage easily of reproductive growth at it, when environmental temperature is lower than 23.5-24.0 ℃, the fertility of photo-thermo-sensitive genic male sterile line begins conversion, can be educated by sterile being converted to, thereby show as the purity that self-fertility influences hybrid rice seeds.
The Rapid identification of hybrid rice seeds purity is that seed quality detects questions of common interest such as department, R﹠D institution, seed production unit always.At present, the evaluation of hybrid rice seeds purity mainly contains several methods such as strange land plantation survey method, isodynamic enzyme survey method and Molecular Identification method.Strange land plantation survey method is method commonly used, but because the plantation of hybrid rice seeds strange land identifies that usually being arranged in Hainan carries out, generally need a paddy growth cycle, namely sample presentation was planted to Hainan, just can be known qualification result in the 4-5 month of next year annual November, qualification result often all lags behind the sale of seed, and this also causes to have every year because of the seed purity problem and causes seed quality dispute event.In the recent period, indoor authenticate technologies such as isodynamic enzyme survey method and Molecular Identification legally constituted authority are relative ripe, use also comparatively general.Indoor survey method qualification time weak point, the relative cost required than strange land plantation survey method also will reduce a lot.
Salt resistance is widely different between rice varieties.The method of paddy rice salt resistance evaluation both at home and abroad is more, mainly contains seed germination evaluation, evaluation in seedling stage, field Screening and Identification, methods such as culturing room's biochemical identification.
Summary of the invention
Yancheng kind tomorrow industry Science and Technology Ltd. finds in practices in several years of screening salt water resistance rice varieties, the female parent two of the double-linear hybrid rice salt two excellent 888 that Jiangsu Inst. of Coastal Area Agricultural Sciences breeds is that the salt resistance ability of photo-thermo-sensitive genic male sterile line salt 582S in the growth of seedling phase reaches 0.5% left and right horizontal, apparently higher than general rice varieties, comprise double-linear hybrid rice salt two excellent 888.Therefore, utilize this principle, salt two excellent 888 seeds are carried out seedling phase growth approval test under 0.5% salt (Nacl) concentration stress conditions, the performance that can coerce salt according to seedling, the self-fruitful seed of its maternal salt 582S in just can identifying between salt two excellent 888 seed farms, thus can be accurately, quick, the practical purity of identifying salt two excellent 888 seeds.
The object of the present invention is to provide that a kind of purity to double-linear hybrid rice salt two excellent 888 seeds is carried out accurately, quick, the practical method that detects.
Technical scheme of the present invention is:
A kind of double-linear hybrid rice salt two excellent 888 seed purity detection methods, its concrete steps are:
(1) conventional presoaking and germinating: select salt two excellent 888 seeds of full seed, neat and consistent, no breakage, no mildew more than 1500 (contrast salt 582S seed is more than 500), according to conventional way carry out disinfection, seed soaking, vernalization, appropriateness is wanted in vernalization, and the grasp seed breaks chest, showing money or valuables one carries unintentionally gets final product.
(2) husky training matrix is prepared: earlier the fine sand after the big or small uniformity sterilization is contained in the culture dish (case) height 1.5-2.0cm; The concentration for preparing toward the middle adding of culture dish (case) is that moistening to fine sand, the suitable seed germination of 0.5% salt (Nacl) solution gets final product, 4 repetitions (containing 1 repetition of contrast), and each repeat request is sowed more than 500.
(3) salt (Nacl) is coerced and cultivated seedling: that will break that chest shows money or valuables one carries unintentionally is examined seed evenly on the sand training matrix of sowing culture dish (case) in, covers with the fine sand of sterilizing then, does not reveal to plant to get final product; After planting culture dish (case) is placed in the climatic cabinate, and temperature keeps 28-30 ℃, and humidity keeps 70-75%; The thin water layer that keeps 0.5% salt (Nacl) concentration in the culture dish behind the 3-5d (case), simultaneously every 12h changes 0.5% salt (Nacl) solution, to reduce the variation of salinity.
(4) growth of seedling is observed record: the back of emerging is to 3-4 leaf phase (after planting about 15d), observe continuously record growth of seedling situation, (showing not, salt tolerant is salt two excellent 888 hybrid F to the seedling quantity that statistics occurs continuously that blade tip takes place to curl, wilting appears in blade, growth slows down symptom
1), and rice shoot quantity identical with contrast under salt (Nacl) stress conditions, that growth is normal (the performance salt tolerant is salt 582S self-fertility seed).
(5) calculating is subjected to sample product purity: being subjected to sample product purity is 3 repetition mean values, is the actual purity of salt two excellent 888 seeds.Be subjected to sample product purity computing formula to be:
Be subjected to sample product purity (%)=(for the total strain number of sample product seedling-Fei Ben kind seedling strain number) ∕ for the total strain number of sample product seedling * 100.
The present invention utilizes according to after years of research and found that the salt resistance of the maternal photo-thermo-sensitive genic male sterile line salt 582S seedling phase of the double-linear hybrid rice salt two excellent 888 that Jiangsu Inst. of Coastal Area Agricultural Sciences breeds is significantly higher than the principle of general rice varieties, under 0.5% salt (Nacl) concentration stress conditions, salt two excellent 888 seeds and salt 582S seed (contrast) are cultivated seedling in the laboratory, emerge the back to 3-4 leaf phase (after planting about 15d), observe record growth of seedling situation, occur blade tip continuously and take place to curl, wilting appears in blade, growth slows down symptom, illustrate that salt tolerant is not salt two excellent 888 hybrid F
1And growing under salt (Nacl) stress conditions normally, illustrating that salt tolerant is its maternal salt 582S self-fertility seed between quaternary salt two excellent 888 seed farms, thereby can calculate the purity of salt two excellent 888 seeds.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
A kind of double-linear hybrid rice salt two excellent 888 seed purity detection methods, its concrete steps are:
(1) presoaking and germinating: select salt two excellent 888 seeds of full seed, neat and consistent, no breakage, no mildew more than 1500 (contrast salt 582S seed is more than 500), with seed with the 0.1%HgCl 10min that sterilizes, use distilled water flushing 4-5 time then, seed soaking, vernalization again, appropriateness is wanted in vernalization, and the grasp seed breaks chest, showing money or valuables one carries unintentionally gets final product.
(2) husky training matrix is prepared: earlier the fine sand after the big or small uniformity sterilization is contained in the culture dish (case) height 1.5-2.0cm; The concentration for preparing toward the middle adding of culture dish (case) is that moistening to fine sand, the suitable seed germination of 0.5% salt (Nacl) solution gets final product, 4 repetitions (containing 1 repetition of contrast), and each repeat request is sowed more than 500.Each repeats and will a plurality of culture dishes (case) can be set according to the size of culture dish (case).Described concentration is that 0.5% salt (Nacl) solution is that to be dissolved in the distilled water of 1000ml by 5gNacl formulated.
(3) salt (Nacl) is coerced and cultivated seedling: that will break that chest shows money or valuables one carries unintentionally is examined seed evenly on the sand training matrix of sowing culture dish (case) in, covers with the fine sand of sterilizing then, does not reveal to plant to get final product; After planting culture dish (case) is placed in the climatic cabinate, and temperature keeps 28-30 ℃, and humidity keeps 70-75%; The thin water layer that keeps 0.5% salt (Nacl) concentration in the culture dish behind the 3-5d (case), simultaneously every 12h changes 0.5% salt (Nacl) solution, to reduce the variation of salinity.
(4) growth of seedling is observed record: the back of emerging is to 3-4 leaf phase (after planting about 15d), observe continuously record growth of seedling situation, (showing not, salt tolerant is salt two excellent 888 hybrid F to the seedling quantity that statistics occurs continuously that blade tip takes place to curl, wilting appears in blade, growth slows down symptom
1), and rice shoot quantity identical with contrast under salt (Nacl) stress conditions, that growth is normal (the performance salt tolerant is salt 582S self-fertility seed).
(5) calculating is subjected to sample product purity: being subjected to sample product purity is 3 repetition mean values, is the actual purity of salt two excellent 888 seeds.Be subjected to sample product purity computing formula to be:
Be subjected to sample product purity (%)=(for the total strain number of sample product seedling-Fei Ben kind seedling strain number) ∕ for the total strain number of sample product seedling * 100.
In the past few years, salt two excellent 888 seed purity assays and field planting evaluation purity result is compared the unanimity of height as a result of the two (seeing Table 1,2,3).
Table 1:2010 utilizes salt resistance to detect salt two excellent 888 seed purities and field planting is identified the purity table of comparisons.
| Sample number into spectrum | Field qualification result (%) | Salt tolerant qualification result (%) |
| 1 | 98.1 | 99.0 |
| 2 | 98.4 | 98.5 |
| 3 | 98.6 | 98.7 |
| 4 | 98·5 | 98·8 |
| 5 | 98.6 | 98.9 |
| 6 | 98·7 | 98·9 |
| 7 | 98.2 | 98.6 |
| 8 | 98.6 | 98.8 |
| 9 | 97·8 | 98·0 |
| 10 | 98.5 | 98.7 |
| 11 | 97.9 | 98.1 |
| 12 | 97.6 | 97.8 |
| 13 | 97.4 | 97.6 |
| 14 | 98.1 | 98.3 |
| 15 | 98.3 | 98.5 |
| 16 | 98·0 | 98·1 |
| 17 | 98.1 | 98.0 |
| 18 | 98.0 | 98.1 |
| 19 | 98.4 | 98.6 |
| 20 | 98.3 | 98.5 |
Table 2:2011 utilizes salt resistance to detect salt two excellent 888 seed purities and field planting is identified the purity table of comparisons.
| Sample number into spectrum | Field qualification result (%) | Salt tolerant qualification result (%) |
| 1 | 98.6 | 98·8 |
| 2 | 98.1 | 98·3 |
| 3 | 97.6 | 97·9 |
| 4 | 97.3 | 97·5 |
| 5 | 98·0 | 98·2 |
| 6 | 98.3 | 98·5 |
| 7 | 97·5 | 97·6 |
| 8 | 97.8 | 98·0 |
| 9 | 97.9 | 98·1 |
| 10 | 96.9 | 97·0 |
| 11 | 97.2 | 97·5 |
| 12 | 97.6 | 97·7 |
| 13 | 97.2 | 97·4 |
| 14 | 97.8 | 98·0 |
| 15 | 98.0 | 98·2 |
| 16 | 98.1 | 98·3 |
| 17 | 97.8 | 98·1 |
| 18 | 97.5 | 97·6 |
| 19 | 97.4 | 97·6 |
| 20 | 97.5 | 97·6 |
| 21 | 98.6 | 98·8 |
Table 3:2012 utilizes salt resistance to detect salt two excellent 888 seed purities and field planting is identified the purity table of comparisons.
| Sample number into spectrum | Field qualification result (%) | Salt tolerant qualification result (%) |
| 1 | 98.6 | 98.6 |
| 2 | 98.5 | 98.6 |
| 3 | 97·5 | 97·7 |
| 4 | 97.3 | 97.5 |
| 5 | 98.3 | 98.3 |
| 6 | 98·0 | 98·0 |
| 7 | 98.0 | 98.1 |
| 8 | 98·4 | 98·6 |
| 9 | 98.1 | 98.2 |
| 10 | 98.4 | 98.5 |
| 11 | 98.2 | 98.3 |
| 12 | 97.5 | 97.6 |
| 13 | 96.9 | 97.1 |
| 14 | 97.5 | 97.6 |
| IS | 97·3 | 97·5 |
| 16 | 97.6 | 97.8 |
| 17 | 96.9 | 97.1 |
| 18 | 96.8 | 97.2 |
| 19 | 98.3 | 98.5 |
| 20 | 98.6 | 98.9 |
| 21 | 97.8 | 97.9 |
| 22 | 97.6 | 97.9 |
| 23 | 96·8 | 97·1 |
| 24 | 97.5 | 97.6 |
| 25 | 97.4 | 97.6 |
Utilized salt resistance to detect salt two excellent 888 seed purities and field planting evaluation purity result in 2010,2011,2012, test through t: the difference of utilizing salt resistance to detect between salt two excellent 888 seed purities and the field planting evaluation purity result in each year is not remarkable, and the purity result's that two kinds of distinct methods detect uniformity is described.Also explanation utilizes the difference of different rice varieties salt resistances not only can identify the authenticity of double-linear hybrid rice salt two excellent 888 and the check of seed purity simultaneously, and with respect to the field planting authentication method, its purity detecting is accurate, quick, practical.
Should be understood that, for those of ordinary skills, can be improved according to the above description or conversion, and all these improvement and conversion all should belong to the protection domain of claims of the present invention.
Claims (1)
1. double-linear hybrid rice salt two excellent 888 seed purity detection methods is characterized in that its concrete steps are:
(1) presoaking and germinating;
(2) husky training matrix is prepared: earlier the fine sand after the big or small uniformity sterilization is contained in the culture dish (case) height 1.5-2.0cm; The concentration for preparing toward the middle adding of culture dish (case) is that moistening to fine sand, the suitable seed germination of 0.5% salt (Nacl) solution gets final product, 4 repetitions (containing 1 repetition of contrast), and each repeat request is sowed more than 500;
(3) salt (Nacl) is coerced and cultivated seedling: that will break that chest shows money or valuables one carries unintentionally is examined seed evenly on the sand training matrix of sowing culture dish (case) in, covers with the fine sand of sterilizing then, does not reveal to plant to get final product; After planting culture dish (case) is placed in the climatic cabinate, and temperature keeps 28-30 ℃, and humidity keeps 70-75%; The thin water layer that keeps 0.5% salt (Nacl) concentration in the culture dish behind the 3-5d (case), simultaneously every 12h changes 0.5% salt (Nacl) solution, to reduce the variation of salinity;
(4) growth of seedling is observed record: the back of emerging is to the 3-4 leaf phase, and continuous observation record growth of seedling situation, statistics occur blade tip continuously seedling quantity curling, that the blade appearance is wilted, growth slows down symptom takes place, and showing not, salt tolerant is salt two excellent 888 hybrid F
1And identical with contrast under condition of salt stress, the normal rice shoot quantity of growth, the performance salt tolerant is salt 582S self-fertility seed;
(5) calculating is subjected to sample product purity: being subjected to sample product purity is 3 repetition mean values, is the actual purity of salt two excellent 888 seeds.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103782900A (en) * | 2014-03-05 | 2014-05-14 | 江苏胜田农业科技发展有限公司 | Method for quickly screening salt-resistant rice variety |
| CN105284564A (en) * | 2015-10-10 | 2016-02-03 | 江苏沿海地区农业科学研究所 | Purity detection method for two-type-line sterile-line seeds Y4-2AB of rape |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997013843A1 (en) * | 1995-10-12 | 1997-04-17 | Cornell Research Foundation, Inc. | Production of water stress or salt stress tolerant transgenic cereal plants |
| CN1922974A (en) * | 2005-09-02 | 2007-03-07 | 广东海洋大学 | Production method for hybrid rice seed with induced salt endurance |
| CN101385438A (en) * | 2008-10-29 | 2009-03-18 | 中国科学院东北地理与农业生态研究所 | Screening crop anti reversibility strain |
| CN102106259A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for improving salt endurance of cereal crops |
| CN102349441A (en) * | 2011-08-05 | 2012-02-15 | 江苏沿海地区农业科学研究所 | Method for breeding coastline salt-resistant paddy |
-
2013
- 2013-06-26 CN CN201310261689.9A patent/CN103329660B/en not_active Expired - Fee Related
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1997013843A1 (en) * | 1995-10-12 | 1997-04-17 | Cornell Research Foundation, Inc. | Production of water stress or salt stress tolerant transgenic cereal plants |
| CN1922974A (en) * | 2005-09-02 | 2007-03-07 | 广东海洋大学 | Production method for hybrid rice seed with induced salt endurance |
| CN101385438A (en) * | 2008-10-29 | 2009-03-18 | 中国科学院东北地理与农业生态研究所 | Screening crop anti reversibility strain |
| CN102106259A (en) * | 2009-12-24 | 2011-06-29 | 上海市农业科学院 | Method for improving salt endurance of cereal crops |
| CN102349441A (en) * | 2011-08-05 | 2012-02-15 | 江苏沿海地区农业科学研究所 | Method for breeding coastline salt-resistant paddy |
Non-Patent Citations (2)
| Title |
|---|
| 柯玉琴 等: "鉴定水稻发芽种子成苗过程中耐盐性的NaCl琼脂固定法", 《植物生理学通讯》 * |
| 王建飞 等: "盐胁迫浓度和胁迫时的温度对水稻耐盐性的影响", 《中国水稻科学》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103782900A (en) * | 2014-03-05 | 2014-05-14 | 江苏胜田农业科技发展有限公司 | Method for quickly screening salt-resistant rice variety |
| CN105284564A (en) * | 2015-10-10 | 2016-02-03 | 江苏沿海地区农业科学研究所 | Purity detection method for two-type-line sterile-line seeds Y4-2AB of rape |
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