CN103329660B - 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, is more particularly a kind of excellent 888 Seed purity test methods of double-linear hybrid rice salt two.
Background technology
Paddy rice is the first cereal crops of China, and China is hybrid rice seeds producing country maximum in the world and country of consumption, and producing highly purified hybrid rice seed is all the basis giving full play to hybrid vigour yield potential.In the recent period, the research and extension of double-linear hybrid rice is that China's Rice Production has made positive contribution, female parent due to double-linear hybrid rice is photo-thermo-sensitive genic male sterile line, photo-thermo-sensitive genic male sterile line enters the impact being easily subject to the environmental condition such as illumination, temperature early stage of reproductive growth at it, when environmental temperature is lower than 23.5-24.0 DEG C, the fertility of photo-thermo-sensitive genic male sterile line starts conversion, can be educated by sterile being converted to, and shows as self-fertility thus affects the purity of hybrid rice seeds.
The Rapid identification of hybrid rice seeds purity is that seed quality detects the questions of common interest such as department, R&D institution, seed produces unit always.At present, the qualification of hybrid rice seeds purity mainly contains the several methods such as Planting in the different location survey method, Identification of Isozyme method and molecular methods.Planting in the different location survey method is the method comparatively commonly used, but carry out because the qualification of hybrid rice seeds Planting in the different location is usually arranged in Hainan, a paddy growth cycle of general needs, 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 and causes seed quality dispute event because of seed purity problem.In the recent period, the indoor authenticate technology relative maturity such as Identification of Isozyme method and Molecular Identification legally constituted authority, apply also comparatively general.Indoor survey method is short compared with the qualification time needed for Planting in the different location survey method, relative cost also will reduce a lot.
Between rice varieties, salt resistance is widely different.The method of domestic and international Salt Resistance of Rice qualification is more, and mainly contain seed germination qualification, seedling stage assay, Field Screening is identified, the methods such as culturing room's biochemical identification.
Summary of the invention
Yancheng Mingtian Seed Industry Technology Co., Ltd. finds in practices in several years of Selection of Salt-Tolerant rice varieties, the female parent two of the double-linear hybrid rice salt two excellent 888 that Jiangsu Inst. of Coastal Area Agricultural Sciences is bred as is that photo-thermo-sensitive genic male sterile line salt 582S reaches 0.5% left and right horizontal at the salt resistance ability of seedling growth stage, apparently higher than general rice varieties, comprise double-linear hybrid rice salt two excellent 888.Therefore, utilize this principle, excellent for salt two 888 seeds are carried out Growth of Seedling approval test under 0.5% salt (Nacl) concentration stress conditions, the performance can coerced salt according to seedling, just can identify the self-fruitful seed of its maternal salt 582S between excellent 888 seed farms of salt two, thus can accurately, the purity of quick, excellent 888 seeds of practical qualification salt two.
The object of the present invention is to provide a kind of purity to excellent 888 seeds of double-linear hybrid rice salt two to carry out accurately, quick, the practical method detected.
Technical scheme of the present invention is:
A kind of excellent 888 Seed purity test methods of double-linear hybrid rice salt two, its concrete steps are:
(1) conventional presoaking and germinating: select full seed, neat and consistent, without damaged, without excellent more than 1500,888 seed of salt two (contrasting more than 500, salt 582S seed) of mildew, conveniently way carry out disinfection, soak seed, vernalization, appropriateness is wanted in vernalization, grasps seed and breaks chest, shows money or valuables one carries unintentionally.
(2) husky training matrix prepares: the fine sand after first being sterilized by size uniformity is contained in culture dish (case), height 1.5-2.0cm; In culture dish (case), add the concentration prepared is moistening to fine sand, the applicable seed germination of 0.5% salt (Nacl) solution, and repeat (containing contrast 1 repetition) 4 times, each repeat request sows more than 500.
(3) salt (Nacl) is coerced and is cultivated seedling: what showed money or valuables one carries unintentionally by broken chest evenly sows in the sand training matrix in culture dish (case) by inspection seed, then covers with the fine sand of sterilizing, does not reveal and plant; To after planting be placed in climatic cabinate by culture dish (case), temperature keeps 28-30 DEG C, and humidity keeps 70-75%; Keep the thin water layer of 0.5% salt (Nacl) concentration after 3-5d in culture dish (case), every 12h changes 0.5% salt (Nacl) solution, to reduce the change of salinity simultaneously.
(4) growth of seedling is observed and is recorded: to 3-4 leaf phase (after planting about 15d) after emerging, Continuous Observation records growth of seedling situation, and statistics occurs that blade tip curling, that wilting appears in blade, growth slows down symptom seedling numbers occurs and (shows not salt tolerant and be the excellent 888 hybrid F of salt two continuously
1), and rice shoot quantity that is identical with contrast under salt (Nacl) stress conditions, that grow normal (show salt tolerant and be salt 582S self-fertility seed).
(5) calculate by inspection sample purity: be subject to inspection sample purity to be repeat mean value 3 times, be the actual purity of excellent 888 seeds of salt two.By sample product purity computing formula be:
By inspection sample purity (%)=(for sample product seedling total strain number-Fei Ben kind seedling strain number) ∕ for total strain number × 100 of sample product seedling.
The present invention utilizes the principle being significantly higher than general rice varieties according to the salt resistance of the maternal photo-thermo-sensitive genic male sterile line salt 582S Seedling Stage that after years of research and found that the double-linear hybrid rice salt two excellent 888 that Jiangsu Inst. of Coastal Area Agricultural Sciences is bred as, under 0.5% salt (Nacl) concentration stress conditions, by excellent for salt two 888 seeds and salt 582S seed (contrast) laboratory culture seedling, to 3-4 leaf phase (after planting about 15d) after emerging, observe and record growth of seedling situation, occur that blade tip occurs continuously curling, wilting appears in blade, growth slows down symptom, illustrate that salt tolerant is not the excellent 888 hybrid F of salt two
1, and under salt (Nacl) stress conditions growth normal, illustrate that salt tolerant is its maternal salt 582S self-fertility seed between excellent 888 seed farms of upper quaternary salt two, thus the purity of excellent 888 seeds of salt two can be calculated.
Embodiment
Below in conjunction with specific embodiment, the present invention is described in detail.
A kind of excellent 888 Seed purity test methods of double-linear hybrid rice salt two, its concrete steps are:
(1) presoaking and germinating: select full seed, neat and consistent, without damaged, without excellent more than 1500,888 seed of salt two (contrasting more than 500, salt 582S seed) of mildew, sterilize seed 0.1%HgCl 10min, then distilled water flushing 4-5 time is used, soak seed again, vernalization, appropriateness is wanted in vernalization, grasps seed and breaks chest, shows money or valuables one carries unintentionally.
(2) husky training matrix prepares: the fine sand after first being sterilized by size uniformity is contained in culture dish (case), height 1.5-2.0cm; In culture dish (case), add the concentration prepared is moistening to fine sand, the applicable seed germination of 0.5% salt (Nacl) solution, and repeat (containing contrast 1 repetition) 4 times, each repeat request sows more than 500.Each repetition will can arrange multiple culture dish (case) according to the size of culture dish (case).To be 0.5% salt (Nacl) solution be described concentration that to be dissolved in the distilled water of 1000ml by 5gNacl formulated.
(3) salt (Nacl) is coerced and is cultivated seedling: what showed money or valuables one carries unintentionally by broken chest evenly sows in the sand training matrix in culture dish (case) by inspection seed, then covers with the fine sand of sterilizing, does not reveal and plant; To after planting be placed in climatic cabinate by culture dish (case), temperature keeps 28-30 DEG C, and humidity keeps 70-75%; Keep the thin water layer of 0.5% salt (Nacl) concentration after 3-5d in culture dish (case), every 12h changes 0.5% salt (Nacl) solution, to reduce the change of salinity simultaneously.
(4) growth of seedling is observed and is recorded: to 3-4 leaf phase (after planting about 15d) after emerging, Continuous Observation records growth of seedling situation, and statistics occurs that blade tip curling, that wilting appears in blade, growth slows down symptom seedling numbers occurs and (shows not salt tolerant and be the excellent 888 hybrid F of salt two continuously
1), and rice shoot quantity that is identical with contrast under salt (Nacl) stress conditions, that grow normal (show salt tolerant and be salt 582S self-fertility seed).
(5) calculate by inspection sample purity: be subject to inspection sample purity to be repeat mean value 3 times, be the actual purity of excellent 888 seeds of salt two.By sample product purity computing formula be:
By inspection sample purity (%)=(for sample product seedling total strain number-Fei Ben kind seedling strain number) ∕ for total strain number × 100 of sample product seedling.
In the past few years, the excellent 888 Seed purity assessment results of salt two and field trapping test purity result compare, the result height consistent (see table 1,2,3) of the two.
Table 1:2010 utilizes salt resistance to detect excellent 888 seed purities of salt two and the field trapping test purity table of comparisons.
| Sample number into spectrum | Field test result (%) | Tolerant salt 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 excellent 888 seed purities of salt two and the field trapping test purity table of comparisons.
| Sample number into spectrum | Field test result (%) | Tolerant salt 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 excellent 888 seed purities of salt two and the field trapping test purity table of comparisons.
| Sample number into spectrum | Field test result (%) | Tolerant salt 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 |
Salt resistance within 2010,2011,2012, is utilized to detect excellent 888 seed purities of salt two and field trapping test purity result, test through t: the difference utilizing salt resistance to detect between excellent 888 seed purities of salt two and field trapping test purity result in each year is not remarkable, and the uniformity of the purity result that two kinds of distinct methods detect is described.Also illustrate utilizes the difference of different rice varieties salt resistance not only can identify the authenticity of double-linear hybrid rice salt two excellent 888 and the inspection of seed purity simultaneously, and relative to field trapping test 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 convert, and all these improve and convert the protection domain that all should belong to claims of the present invention.
Claims (1)
1. the excellent 888 Seed purity test methods of double-linear hybrid rice salt two, it is characterized in that, its concrete steps are:
(1) presoaking and germinating: select full seed, neat and consistent, without damaged, without excellent 888 seeds of salt two of mildew, carry out disinfection, soak seed, vernalization, appropriateness is wanted in vernalization, and grasp seed breaks chest, shows money or valuables one carries unintentionally;
(2) husky training matrix prepares: the fine sand after first being sterilized by size uniformity is contained in culture dish, height 1.5-2.0cm; In culture dish, add the mass concentration prepared is moistening to fine sand, the applicable seed germination of 0.5%Nacl solution, and repeat for 4 times, repeat containing a contrast, each repeat request sows more than 500;
(3) salt stress cultivates seedling: what showed money or valuables one carries unintentionally by broken chest evenly sows in the sand training matrix in culture dish by inspection seed, then covers with the fine sand of sterilizing, does not reveal and plant; To after planting be placed in climatic cabinate by culture dish, temperature keeps 28-30 DEG C, and humidity keeps 70-75%; Keep the thin water layer of mass concentration 0.5%Nacl after 3-5d in culture dish, every 12h changes a 0.5%Nacl solution, to reduce the change of salinity simultaneously;
(4) growth of seedling is observed and is recorded: to the 3-4 leaf phase after emerging, Continuous Observation records growth of seedling situation, statistics occurs that curling, that wilting appears in blade, growth slows down symptom seedling numbers occurs blade tip continuously, shows not salt tolerant and is excellent 888 Hybrids F1 of salt two; And under condition of salt stress with contrast identical, grow normal rice shoot quantity, performance salt tolerant be salt 582S self-fertility seed;
(5) calculate by inspection sample purity: be subject to inspection sample purity to be repeat mean value 3 times, be the actual purity of excellent 888 seeds of salt two.
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| 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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| CN100566552C (en) * | 2005-09-02 | 2009-12-09 | 广东海洋大学 | A kind of production method for hybrid rice seed with the salt resistance of inducing |
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