GB/T 29371.5-2012 English PDF (GBT29371.5-2012)

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GB/T 29371.5-2012: Technical rules for seed production system of two-line hybrid rice -- Part 5: Technical rules for identification of variety purity and male sterility of MS line
GB/T 29371.5-2012
GB
ICS 65.020.20
B 21
NATIONAL STANDARD
OF THE PEOPLE’S REPUBLIC OF CHINA
Technical rules for seed production system of two-line hybrid
rice - Part 5. Technical rules for identification of variety purity
and male sterility of MS line
ISSUED ON. DECEMBER 31, 2012
IMPLEMENTED ON. JULY 1, 2013
Issued by. General Administration of Quality Supervision, Inspection and
Quarantine of the People's Republic of China;
Standardization Administration Committee of the People's
Republic of China.
Table of Contents
Foreword ... 3 
1  Scope ... 4 
2  Normative references ... 4 
3  Fertility monitoring of sterile line ... 4 
4  Identification of seed purity planting ... 8 
Foreword
GB/T 29371 Technical rules for seed producing system of two-line hybrid rice consists of
five parts, as follows.
— Part 1. Terminology;
— Part 2. Technical rules for foundation seed production of photo/thermo-sensitive genic
male sterile line;
— Part 3. Technical rules for seed multiplication of photo/thermo-sensitive genic male
sterile line;
— Part 4. Technical rules for F1 seed production;
— Part 5. Technical rules for identification of variety purity and male sterility of MS line.
This is Part 5 of GB/T 29371.
This Part is drafted according to the rules specified in GB/T 1.1-2009.
This Part was proposed by Ministry of Agriculture of the People’s Republic of China.
This Part shall be under the jurisdiction of National Technical Committee (SAC/TC 37) on
Crop Seed of Standardization Administration of China.
The responsible drafting organizations. Hunan Hybrid Rice Research Center, Hunan
Longping Seed Industry Co., Ltd, Hunan Agricultural University, Hunan Seed
Administration, Anhui Academy of Agriculture, Jiangsu Academy of Agriculture, and
Guangdong Academy of Agriculture.
The chief drafting staffs of this standard. Zhou Chengshuo, Liu Aimin, Li Wenxiang, Xiao
Cenglin, Wang Shouhai, Lyu Chuangen, Wang Feng, Liao Cuimeng, Liu Jianbing, and
Ouyang Aihui.
Technical rules for seed production system of two-line hybrid rice –
Part 5. Technical rules for identification of variety purity and male sterility
of MS line
1 Scope
This Part of GB/T 29371 specifies the methods and procedures for monitoring the critical
temperature for inducing male sterility of the photo/thermo-sensitive genic male sterile line
of rice of the high temperature inducing male sterility line, and identifying the purity of male
sterile seeds and hybrid seeds planting.
This Part is applicable to monitoring of the determined or identified critical temperature for
inducing male sterility of photo/thermo-sensitive genic male sterile line of the high
temperature inducing male sterility line, and identification of the purity of male sterile
seeds and hybrid seeds planting.
2 Normative references
The articles contained in the following documents have become part of this standard when
they are quoted herein. For the dated documents so quoted, all the modifications
(Including all corrections) or revisions made thereafter shall be applicable to this Standard.
GB 4404.1-2008 Seed of food crops - Part 1. Cereals
GB/T 3543.2-1995 Rules for agricultural seed testing-Sampling
3 Fertility monitoring of sterile line
3.1 Monitoring of phytotron
3.1.1 Monitoring materials and requirements
3.1.1.1 Monitoring materials
The photo/thermo-sensitive genic male sterile line of rice that passes the examination or
technical identification by the provincial or above-level Crop Variety Approval Committee,
and being used in production.
3.1.1.2 Material requirements
Identification materials require 300g-500g of seeds; germination rate ≥80%; and purity
≥99.5%. And provide the breed standard when the sterile line passed the approval or
identification.
3.1.1.3 Sampling requirements
It shall execute the provisions of GB/T 3543.2-1995.
sterile line for identification). See Table 1 for temperature of each period.
Table 1 Temperature and light setting in phytotron with daily average temperature
of 23.5°C or 23.0°C
Temperature
Sunshine Time
Daily average
temperature
23.5°C
Daily average
temperature
23.0°C
06. 01-10. 00 23.5°C 23.0°C Light intensity ≥1 × 104 lx
20.00 - 6.00 is deemed as dark period (14.0h)
19.30 - 6.00 is deemed as dark period (13.5h)
18.30 - 6.00 is deemed as dark period (12.5h)
10. 01-18. 00 26.5°C 26.0°C
18. 01-22. 00 23.5°C 23.0°C
22. 01-06. 00 20.5°C 20.0°C
3.1.3.5 Temperature and light setting during transitional period
2d before and after the processing, if the natural daily average temperature is ≥ 30°C or ≤
25°C, then the processing materials shall be transited for 2d under the conditions –
day-time temperature of 28°C, night-time temperature of 24°C, and light intensity of
1.0×104 lx, and then move them into the phytotron or place them in a natural conditions.
3.1.4 Fertility observation
3.1.4.1 Observed object
When the process in phytotron is completed, tag and mark the rice spikes of which the
distance between flag-leaf and top-second leaf is 0 cm - 2 cm.
3.1.4.2 Observation method
When the marked spikes grow into the initial heading stage, every morning take the
glumous flowers that will be flowering on that day (anthers length stretching to two-thirds
of glumous flower) to conduct pollen observation under microscope. Randomly pick 5-6
glumous flowers from each ear; their anther is made into 1 observing piece. Use I2-KI
solution to dye the pollens and press into piece. Under the optical microscope, amplify
100 times to inspect. Choose 3 representative views for each piece. Pollen particles of
each view shall be more than 50. Observe and count the numbers of different type of
pollens of 3 views (non-pollen abortion type, typical abortion type, spherical abortion type,
stained abortion type, and normal). Calculate the percentage of different type of pollens in
each piece. If stained pollen is found in an observation piece, then it is deemed as male
sterility alteration.
3.1.4.3 Times of observations
Observation under microscope shall be conducted once a day, until the marked spikes
end the flowering.
3.1.4.4 Pollen classification
3.2.4 Fertility examination
3.2.4.1 Pollen observation under microscope
From the beginning of flowering to the latest normal heading, observation under
microscope shall be conducted once every 2d-3d. If there are staining pollens found
during this period, the pollen observation under microscope shall be conducted every day,
from that day until two days after the day without staining pollens.
3.2.4.2 Examination of self-seeding rate
Randomly select 10 single-plants from the seeding period when staining pollens are found
in observation under microscope. Completely isolate them to cultivate. 20d-25d after full
heading, examine the self-seed rate.
3.2.5 Investigation for sterile line group uniformity
Select two completely sterile seeding periods for each sterile line. Investigate the growth
period, plant height, plant leaf and ear grain type, color, and other characters.
3.2.6 Comprehensive evaluation
3.2.6.1 Agronomic characters
According to the monitored materials’ field’s fertile property, plant morphological
characteristic, ear grain shape, outcross characteristics, resistance, and other
performances, evaluate the stability and conformity of monitored materials group, in
combination with breed introduction.
3.2.6.2 Critical temperat...