Crops ›› 2023, Vol. 39 ›› Issue (4): 71-76.doi: 10.16035/j.issn.1001-7283.2023.04.011

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Comparison ofPhenotype and Marker Detection in Seed Purity of Thermo-Photo Sensitive Two-Line WheatHybrids

Li Hongsheng1(), Li Shaoxiang1(), Yang Zhonghui1, Yang Jiali2, Liu Kun1, Xiong Shian3, Li Fuqian1, Guo Hui1, Yang Mujun1()   

  1. 1Institute of Food Crops, Yunnan Academy of Agricultural Sciences, Kunming 650205, Yunnan, China
    2College of Agriculture, Yunnan Agricultural University, Kunming 650201, Yunnan, China
    3Seed Management Station of Zhenxiong County, Zhaotong 657200, Yunnan, China
  • Received:2022-03-21 Revised:2022-04-06 Online:2023-08-15 Published:2023-08-15

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Abstract:

The purity identification of hybrid seeds is one of key steps in the production and application of thermo-photo sensitive wheat hybrids. At present, seed purity is determined mainly by field phenotypic identification of planted hybrids in the field, which takes a long time.The purity of two elite combinations K64S/20Y4-5 and K64S/MR1238 were tested by phenotypic identification in field and SSR fluoresent marker screening. Meanwhile, the results of identification of two ways were compared. The results showed that the purity of two hybrids were 98.84% and 97.93% respectively through phenotypic identification in field. There were three pairs of primers (barc164, gwm161 and gwm610) of eight pairs of primers showed polymorphism between parents. The purities of two hybrids were 97.73% and 97.21% using primer of barc164, respectively, slightly lower than those of phenotypic identification in field, but there was no significantly difference between two methods. In addition, SSR fluoresent marker could not only distinguish seeds including self-cross single plant of sterile line and mechanical mixture of male parent, but also accurately identify pseudo-hybrids caused by impure male parents and outcrossing with foreign pollens in hybrid seed production. Therefore, SSR fluorescent markers could be replaced phenotypic identification for quickly testing the purity of hybrids.

Key words: Wheat, Thermo-photo sensitive genic male sterile line, Hybrid, Purity, SSR

Table 1

Primersequences of SSR markers"

引物名称Primer name 染色体分布Chromosome location 引物序列（5′-3′）The sequence of primer(5′-3′)
barc324 3A F：CCAATTCTGCCCATAGGTGA
R：GAGGAAATAAGATTCAGCCAACTG
barc164 3B F：TGCAAACTAATCACCAGCGTAA
R：CGCTTTCTAAAACTGTTCGGGATTTCTAA
cfa2028 7A F：TGGGTATGAAAGGCTGAAGG
R：ATCGCGACTATTCAACGCTT
cfa2123 7A F：CGGTCTTTGTTTGCTCTAAACC
R：ACCGGCCATCTATGATGAAG
gwm155 3A F：CAATCATTTCCCCCTCCC
R：AATCATTGGAAATCCATATGCC
gwm161 3D F：GATCGAGTGATGGCAGATGG
R：TGTGAATTACTTGGACGTGG
gwm304 5A F：AGGAAACAGAAATATCGCGG
R：AGGACTGTGGGGAATGAATG
gwm610 4A F：AGGAAACAGAAATATCGCGG
R：AGGACTGTGGGGAATGAATG

Table 2

Phenotyping identificationin plant height ofhybrids"

杂交种
Hybrid		 鉴定株数
Number of
tested plants		 表型“1”株数
Number of plants
with phenotype “1”		 表型“2”株数
Number of plants
with phenotype “2”		 表型“3”株数
Number of plants
with phenotype “3”		 纯度
Purity
(%)
K64S/MR1238 1467 8 9 1450 98.84
K64S/20Y4-5 1448 23 7 1418 97.93

Table 3

Polymorphic primer screening between parents"

亲本Parent barc164 barc324 cfa2028 cfa2123 gwm155 gwm161 gwm304 gwm610
20Y4-5 211 258 275 270 165 192 222 189
MR1238 199 264 277 270 147 195 222 189
K64S 208 264 277 270 147 174 222 193

Fig.1

The peak spectrum of polymorphic primers of three parents"

Table 4

Comparison of the purity of 574 hybrids tested by phenotype and marker identification"

组合
Combination		 鉴定数
Number
of tested
plants		 表型“1”
株数
Number of
plants with
phenotype “1”		 表型“2”
株数
Number of
plants with
phenotype “2”		 表型“3”
株数
Number of
plants with
phenotype “3”		 表型纯度
Purity
identifiedby
phenotype
(%)		 母本基因
型数量
Number of
femaleparent
genotypes		 父本基因
型数量
Number of
male parent
genotypes		 杂合基因型
Heterozygous
genotype		 其他基因型
Number of
other
genotypes		 标记纯度
Purity
identified
by the
marker(%)
K64S/MR1238 574 4 5 565 98.43 4 5 561 4 97.73
K64S/20Y4-5 574 9 4 561 97.74 9 4 558 3 97.21
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