This standard specifies the method for identifying the fertility of light- and thermo-sensitive male sterile lines of rice in artificial climate chambers and in the field. This standard is applicable to the identification of the fertility of light- and thermo-sensitive male sterile lines of rice. NY/T 1215-2006 Procedure for identifying the fertility of light- and thermo-sensitive male sterile lines of rice NY/T1215-2006 Standard download decompression password: www.bzxz.net
This standard specifies the method for identifying the fertility of light- and thermo-sensitive male sterile lines of rice in artificial climate chambers and in the field. This standard is applicable to the identification of the fertility of light- and thermo-sensitive male sterile lines of rice.

ICS 65.020.20
Agricultural Industry Standard of the People's Republic of China
NY/T1215-2006
Fertility evaluation protocol of environmental sensitivc genic malesterile (EGMS) ine of rice
Published on December 6, 2006
Implemented from February 1, 2007
Ministry of Agriculture of the People's Republic of China
Appendix A of this standard is the normative list.
NY/T1215--2006
This standard is proposed and managed by the Ministry of Agriculture of the People's Republic of China. The main drafting units of this standard are: Quality Supervision, Inspection and Testing Center for Rice and Its Products, National Key Laboratory of Rice Biology, and National Rice Research Institute of the Ministry of Agriculture.
The main drafters of this standard are: Sun Quanxiu, Shi Yonghong, Chen Neng, Suo Huabei, Shi Yongfeng, Wei Xinghua, Hu Guocheng, Xu Li, 1 Scope
Procedure for identification of fertility of light-sensitive and thermosensitive male sterile lines of rice NY/T1215—2006
This standard specifies the method for identification of fertility of light-sensitive and thermosensitive male sterile lines of rice in artificial climate chambers and chambers. This standard is applicable to the identification of fertility of light-sensitive and thermosensitive male sterile lines of rice 2 Terms and definitions
The following terms and definitions apply to this standard. 2.1
Envirommental sesitive genic male sterile (EGMS) line of rice is a fertile male sterile line that is sterile under long-day and/or high-temperature conditions and male-fertile under short-day and/or low-temperature conditions. Usually represented by S, there are many types, which can also be collectively referred to as environmentally sensitive male sterile lines: 2.2
Environmentally sensitive male sterilityEnvironmentally sensitive male sterilityThe degree to which the male gamete development is positive under long-term illumination and/or high temperature conditions, and the degree to which the male gamete development is blocked, resulting in a decrease in the plant fruiting rate, is a key indicator for determining seed production purity. It is expressed by the microscopic pollen sterility and the bagged white cross fruiting rate (i.e., small-scale sterility).
Fertilitity
The degree to which the male gamete development is stopped under short-term illumination and/or suitable low temperature conditions, resulting in the plant fruiting rate, is a key indicator for determining seed yield. It is expressed by the bagged white cross fruiting rate. It is also called fertility. 2.4
Degree of malc sterility is an indicator to measure the degree of sterility of the most sterile line, expressed by the sterility rate of self-pollination and the sterility of pollen. 2.5
Ratio of self-fertility is the fruiting rate of white flower pollination.
Ratio of seif-sterility is the sterility rate of white flower pollination, also known as spikelet sterility. 3 Requirements
The photosensitive nuclear sterile lines that have passed the identification shall be re-inspected according to this standard every year. 4 Fertility evaluation
4.1 Evaluation of the identification results of artificial climate chamber
The evaluation of the light and temperature response characteristics of the fertility conversion of the photosensitive and thermosensitive male sterile lines of rice is divided into two parts: sterility and fertility (fertility). With sterility as a prerequisite, the sterile lines with good sterility shall be further evaluated for fertility. The evaluation of sterility and fertility is based on the self-pollination rate as the main indicator. The standards are shown in Tables 1 and 2. 1
NY/T1215—2006
Table 1 Evaluation standards for the results of light-temperature ecological sterility identification in artificial climate box Small sugar individual fertility
Average cross-fertility rate
Note: Light length is 13.55 or 14.5h
Single socks Minimum simple
Pollen individual fertility
Average pollen sterility
Table 2 Evaluation standards for the results of light-temperature ecological sterility identification in artificial climate box Bag level
:1: Light length is 12.55 or 11.51
Note 2: Temperature is 24℃ or 23℃
Self-pollination rate
5.0--10.0
10.0--20 .0
Single trait: %
Minimum value of single plant
Unit: %
For all sterile lines that meet the requirements of Table 1 at the same time, the data of spikelet sterility were subjected to variance analysis after sin1/transformation. The sterile lines that achieved fertility conversion with significant photoperiod effect and photoeffect but no significant temperature effect were photoperiod-sensitive sterile lines; the sterile lines that achieved fertility conversion with significant temperature effect but no significant photoperiod effect, and significant or no significant photoperiod effect were temperature-sensitive sterile lines; the sterile lines that achieved fertility conversion with no significant photoperiod effect and temperature effect but only significant phototemperature interaction effect were phototemperature interaction sterile lines; the others were non-phototemperature-sensitive male sterile lines
4.2 Identification plot inspection and evaluation
1) The agronomic traits of the population of more than 1000 plants were uniform and effective. 2) Individual fertility rate is 100%, microscopic pollen sterility is 99.5%, and the maximum fertility of a single plant is 3%; the sterility of the small stable seedlings is 99.5%, and the maximum fertility of a single plant is 1%.
The sterility period of the light- and thermo-sensitive male sterile system is long and stable. Under normal natural planting density, the stable sterility period can last for more than 3 days, and the natural fruit setting rate of the period can reach 30%.
5 Inspection method
5.1 Artificial climate box inspection
5.1.1 Sowing and transplanting
The seeds of the light- and thermo-sensitive male sterile system tested are uniformly sown in the identification nursery after sterilization. When the plants grow to the 5-leaf stage, select the plants with normal and consistent growth and put them in the container: spray the pesticide before entering the box to prevent and control diseases and insect pests. The control (the conventional varieties are used as fertile controls, and the sterile lines with stable fertility and annual yield rate are used as sterile controls) are planted at the same time.
5.1.2 Short-term treatment
The materials with photosensitive development are treated with short-term treatment for 10 days at the 5-leaf stage. The treatment conditions are: light 10h (7:00·17:00), humidity 28℃; spray treatment 14h (17:00-7:00), temperature 25℃. After 10 days of treatment, the materials with insensitive development do not need this treatment. 2
5.1.3 Light and temperature treatment
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When the main ear of the sterile line plants enters the stage of differentiation of the primordium of the pistils and stamens, they are put into the box for light and temperature treatment. For each treatment of 10 plants of each light and temperature male sterile line, the temperature in the climate box should be uniform and consistent, with a fluctuation range of +1.3℃ of the set temperature; the light intensity is greater than 30,000 x, and the light quality and wind speed should be able to ensure the normal growth and development of rice. The temperature in the box is a variable temperature with a period of 24h, and the daily temperature difference is 8. Water is used for irrigation in the climate box. The settings of the light cycle, temperature and relative humidity of the daylight artificial climate box are shown in Appendix A. The water is diluted when the ears begin to grow after 25 days of treatment. Two ears are taken from each plant and bagged, and the heading date is recorded. When bagging, take 1 of the top 1.3.4.5, 6, and more than 5 florets, and fix them with Carnoy's fixative (95% ethanol: ethylenediamine 3:1) for inspection. The inspected plants are taken out of the box after 5-7 days of incubation. After taking them out, it is necessary to ensure that the plants grow under normal conditions. 5.1.4 Check the fruit set rate (F) of the bagged white cross when mature. The standard for full grains is the formation of starch. Grains infected with rice smut or rice mountain disease are full grains. Mark the hybrids and calculate the total number of hybrids (see Figure 2) and the hybrid rate (1): Calculate the total fruit set rate of the white cross: bzxZ.net
--the number of full grains of a single white cross;
N--the total number of self-pollination grains in a single bag.
n2×100
I--hybrid rate;
n2…total number of self-pollination plants;
N--the total number of participants in the test of this strain.
When checking the fertility of pollen, check the 3 spikelets in 5.1.3 for each plant. Each flower should be examined in 2 fields of view. About 100 pollen grains should be examined in each field of view. Use 1% KI solution to stain them, and classify them into four types: black staining (fertile), staining failure, round failure, and typical failure. Calculate the pollen sterility (P), the calculation formula:
Where:
Pollen sterility;
The number of sterile pollen examined per plant (the sum of staining failure, round failure, and typical failure); The total number of pollen examined per plant under a microscope,
5.1.5 Data report
a) Calculate the average value and standard deviation of the white cross fruiting rate of each site (after removing the impure plants) according to the light- and temperature-sensitive male sterile lines. b) Report the maximum and minimum values ​​of each treatment, c) Report the total number of sterile impure plants and impure plants. d) Report the pollen sterility.
5.2 Identification and testing
5.2.1 Sterility test
5.2.1.1 Planting
Identification of sex: Plant more than 1000 light- and temperature-sensitive sterile lines in enclosures. When withdrawing, isolate and randomly bag 300 ears. Each plant is limited to 1 ear 3
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5.2.1.2 Inspection
a) Check the sterile plant rate: Check the actual number of plants in the group, which shall not be less than 1000 palms, and there shall be no missing plants in the field, and the number of mixed plants shall be counted. Randomly take 200 bagged ears, and examine the fruiting rate of each household indoors. b) Check the sterility of pollen: Fertility identification: Randomly take 100 ears at heading, and quickly check the pollen fertility under a microscope. The method is to examine the mixed pollen of three spikelets under a microscope for each ear, and count the pollen according to the two categories of black staining (fertile), staining abortion, round abortion, and typical abortion (sterile): 5.2.2 Inspection of continuous sterility period
5.2.2.1 Staged sowing
The first and last stages of staged sowing are respectively 15 days before the earliest sowing date and 15 days after the latest sowing date of the local suitable sowing date of rice, and the interval between the two sowing dates shall not exceed 1.5 days. The seedlings shall be transplanted and planted in a single plant, with no less than 24 plants in each stage. Routine management. 5.2.2.2 Fertility identification
From the month of rice heading in the first stage of sowing to 10 days after the local safe heading date, one ear of heading on the same day shall be randomly taken from each of the three plants of the same period every 1 day, bagged and marked. In order to ensure the connection between the sampling materials of the two adjacent stages, at least 2 simultaneous samplings shall be conducted. 25-30 days after bagging, check the white cross fruiting rate. 5.2.2.3 Calculation of continuous sterility period
The calculation method of continuous sterility period is that the average white cross fruiting rate of 3 ears in the same period is lower than (.5% (single plant value is less than 1%), which is sterility, and the longest number of days of continuous sterility is the number of sterile people in the sterile line. From the differentiation of rice seedlings in the first sowing period to 10 days after the local full ear stage, record the local meteorological data daily, 5.2.3 Data report
) Report the average and standard deviation of the white cross fruiting rate of light-sensitive and temperature-sensitive nuclear sterile lines (such as hybrid plants), h) Report the average and standard deviation of pollen sterility (excluding hybrid plants). c) Report continuous sterility period.
A.1 The light and temperature settings for artificial climate box treatment are shown in Table A1: Appendix A
(Normative Appendix]
Table A1 Light and temperature conditions for artificial climate box treatment Photoperiod
Daily weighted average humidity
And mud degree
Description: Photoperiod humidity: 14.5h: 5:00~19:3013.5up: 5:30~19:00
12.5h: 6:00--18:30
11.5hif:30 ~ 18:00
Daily weighted average temperature, see Table 42:
Three plate temperature
5:00-8:00
8:00--11:00
11:00~16:00
16:00 ~ 20:00
20:00--2:00
2:00~5:00
Table A2 Daily weighted average temperature
NY/T 1215--2006
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