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Acta Agronomica Sinica ›› 2022, Vol. 48 ›› Issue (10): 2654-2662.doi: 10.3724/SP.J.1006.2022.11074

• RESEARCH NOTES • Previous Articles     Next Articles

QTL mapping for salt tolerance in wheat line CH7034

ZHANG Xiao-Wen1(), LI Shi-Jiao1, ZHANG Xiao-Jun2, LI Xin2, YANG Zu-Jun3, ZHANG Shu-Wei2, CHEN Fang2, CHANG Li-Fang2, GUO Hui-Juan2, CHANG Zhi-Jian2,*(), QIAO Lin-Yi2,*()   

  1. 1College of Life Science, Shanxi University, Taiyuan 030006, Shanxi, China
    2College of Agriculture, Shanxi Agricultural University / Shanxi Key Laboratory of Crop Genetics and Molecular Improvement / State Key Laboratory of Sustainable Dryland Agriculture (in preparation), Taiyuan 030031, Shanxi, China
    3College of Life Science and Technology, University of Electronic Science and Technology of China, Chengdu 610054, Sichuan, China
  • Received:2021-08-22 Accepted:2022-01-06 Online:2022-10-12 Published:2022-02-24
  • Contact: CHANG Zhi-Jian,QIAO Lin-Yi E-mail:zxw18435139058@126.com;wrczj@126.com;qiaoly1988@126.com
  • Supported by:
    Shanxi Scholarship Council of China(2021-070);Shanxi Agricultural University Research Project for Doctor(2021BQ39);State Key Laboratory of Sustainable Dryland Agriculture (in preparation) of Shanxi Agricultural University(202002-3)

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Abstract:

The identification of wheat salt-tolerant germplasm is of great significance for making full use of saline land and ensuring food security. CH7034 is a wheat salt-tolerant line bred by our laboratory. In order to clarify the genetic rules and control loci for salt tolerance, QTL analysis was conducted using a recombinant inbred lines population crossed by CH7034 and salt-sensitive variety SY95-71. Based on the SNP microarray data and salt injury index, six QTL were detected on chromosomes 2A, 2D, 4B, and 5A, including QSI.sxau_2A, QSI.sxau_2D, QSI.sxau_4B.1, QSI.sxau_4B.2, QSI.sxau_5A.1, and QSI.sxau_5A.2. Among them, QSI.sxau_5A.1 was detected in all three salt stress experiments and had the highest phenotypic variance explained (15.73%-20.18%), which was different from other salt-tolerance loci reported on chromosome 5AL. Seven SSR markers were developed and integrated in the QSI.sxau_5A.1 interval, and the peak of LOD was further determined at SSR-D1. Based on the transcriptome database, 12 high-confidence genes in response to salt stress were screened from the QSI.sxau_5A.1 section. These results lay the foundation for the fine mapping and even cloning of the salt tolerance loci of CH7034, and provide new germplasm and molecular markers for the selection and breeding of wheat salt-tolerant varieties.

Key words: wheat, salt stress, salt injury index, SNP marker, QTL mapping

Table 1

SSR molecular markers used in this study"

标记名称
Marker name		 上游引物
Forward sequence (5'-3')		 下游引物
Reverse sequence (5'-3')		 产物长度
Product size (bp)		 退火温度
Annealing
temperature (℃)
SSR-U7 GGTCATGTTGGAATGTTGATCAAG GCTTTTATGTTTCCAACTCTAGTCAA 127 58
SSR-D28 GGGAAATCCCAACGCCTG GAGTCATCATGAAAGAAAGCATAAAT 144 58
SSR-D5 GCCCACGTTTCCTCTTAATG TCATCGAACATGATGCACTATC 94 58
SSR-D1 GAGAATAAATCCACTCTCCATGC CGATATGTTCCTGTCTCCTTGT 180 58
SSR-D4 CCGTGTAGTAGAACACTGCATGT GCTCGAGCTGTAGCCCAG 193 54
SSR-U2 AAGCAGAATCATGGAAGGGAC CACTTATTCAAGTCATTAAGCATTCAAT 102 58
SSR-U5 CAGTATCCATCTTTGCCATTCAG CTCCTACCTCGACGACAC 116 58
cslinkNax2 TCTCCATCATTCAACATCAATCG TGTAGCTCGTCGGGGTGTGTTGC 170 58
Xgwm304 AGGAAACAGAAATATCGCGG AGGACTGTGGGGAATGAATG 176 58
Xbarc144 GCGTTTTAGGTGGACGACATAGATAGA GCGCCACGGGCATTTCTCATAC 191 58
Xwmc110 GCAGATGAGTTGAGTTGGATTG GTACTTGGAAACTGTGTTTGGG 127 58

Fig. 1

CH7034 and SY95-71 treated with 250 mmol L-1 NaCl for seven days"

Table 2

Salt injury index of CH7034 and SY95-71 under NaCl stress"

材料
Material		 总株数
Total number of plants		 盐害级别及株数 Salt injury level and number of plants 盐害指数
Salt injury index (%)
0 1 2 3 4
CH7034-1 16 7 8 1 32.5
CH7034-2 16 8 6 2 32.5
CH7034-3 16 8 8 30.0
平均值 Mean 31.7
SY97-71-1 16 2 14 77.5
SY97-71-2 16 3 13 76.3
SY97-71-3 16 2 14 77.5
平均值 Mean 77.1

Fig. 2

Phenotypic distribution of salt injury index in the CH7034/SY95-71 RIL population"

Fig. 3

Genetic length of molecular map constructed by CH7034/SY95-71 RIL population"

Table 3

Mapping of salt-tolerant QTL of CH7034/SY75-71 RILs at seedling stage"

试验编号
Test number		 数量性状位点
QTL		 染色体
Chr.		 图谱位置
Position (cM)		 标记区间
Marker region		 LOD值
LOD score		 表型变异解释率
PVE (%)		 加性效应
Add.
SI-1 QSI.sxau_2D 2D 0 992489-992489 3.57 5.60 -2.42
QSI.sxau_4B.1 4B 112 1265612-3954524 6.38 10.50 -3.31
QSI.sxau_5A.1 5A 73 1049519-4911066 6.63 15.73 -3.81
SI-2 QSI.sxau_2A 2A 9 3532866-1090021 2.96 5.24 -2.71
QSI.sxau_4B.2 4B 43 1862515-1059340 3.41 6.36 -2.96
QSI.sxau_5A.1 5A 71 1049519-1215538 10.29 20.18 -5.49
SI-3 QSI.sxau_5A.1 5A 71 1049519-1215538 10.15 17.62 -4.50
QSI.sxau_5A.2 5A 116 1125437-1228444 4.27 15.73 -3.09

Fig. 4

QSI.sxau-5A.1 section on genetic map (a), genomic map (b), and the LOD map added with SSR markers (c) Marker with asterisk (*) indicates that it is located below the LOD peak."

Fig. 5

Amplification of linkage markers of QSI.sxau-5A and reported salt-tolerance gene/QTL in CH7034, SY75-71, and their RILs a, b: linkage markers SSR-U2 and SSR-U5 of QSI.sxau-5A; c: linkage marker cslinkNax2 of TmHKT1:5-A; d: linkage marker Xbarc144 of the salt tolerance locus of Shanrong 3. M: DNA ladder marker; CH: CH7034; SY: SY95-71; T1-T6: salt-tolerant lines with QSI.sxau-5A; S1-S6: salt-sensitive lines without QSI.sxau-5A."

Table 4

Salt tolerance genes and QTLs on chromosome 5AL reported in wheat"

基因/QTL
Gene/QTL		 参考文献
Reference		 耐盐表型
Phenotype		 来源
Source		 连锁标记 Linkage marker
标记名称
Name		 亲本多态性
Polymorphism
in parents		 与表型连锁Linked to
phenotype
TmHKT1;5-A [6,7] 叶片K+/Na+
K+/Na+ ratio of leaf		 Triticum durum cslinkNax2 + -
QTL-SR3 [9] 叶片盐害级别
Salt injury level of leaf		 wheat Xgwm304 - -
QTL-SR3 [10] 叶片盐害级别
Salt injury level of leaf		 wheat Xbarc144 + -
qNL5, qLL5 [8] 叶数, 叶长
Leaf number, leaf length		 Triticum durum Xwmc110 - -
QHAI.5A-2 [11] 盐碱地收获指数
Harvest index in salt land		 wheat wPt-3334
QG(1-5).asl-5A [12] 相对生长率
Relative growth rate (RGR) salt/RGR control		 wheat TP14539

Table 5

Salt stress response genes in QSI.sxau-5A section selected based on transcriptome database"

差异表达基因
DEGs		 基因组位置
Genomic location		 差异倍数
log2 (Fold Change)		 响应类别
Response type		 编码蛋白注释
Protein-encoding annotation
TraesCS5A01G382800 580799964-580801653 1.71 上调
Upregulate		 钙调素相关钙感应蛋白
Calmodulin-related calcium sensor protein
TraesCS5A01G382900 580803022-580805966 -1.44 下调
Downregulate		 ATP硫酸化酶
ATP sulfurylase
TraesCS5A01G386500 584049719-584058870 -1.08 下调
Downregulate		 肌球蛋白
Myosin protein
TraesCS5A01G388300 585018687-585021441 1.26 上调
Upregulate		 β-葡萄糖苷酶前体
β-glucosidase precursor
TraesCS5A01G389000 585232865-585236702 1.26 上调
Upregulate		 β-葡萄糖苷酶前体
β-glucosidase precursor
TraesCS5A01G390200 585488957-585491921 1.01 上调
Upregulate		 胡豆苷合酶
Strictosidine synthase
TraesCS5A01G392100 588416260-588416877 4.68 上调
Upregulate		 无
None
TraesCS5A01G393900 589311576-589313132 2.25 上调
Upregulate		 UDP-葡萄糖基转移酶
UDP-glucosyl transferase
TraesCS5A01G394500 589584058-589585518 1.38 上调
Upregulate		 UDP-葡萄糖基转移酶
UDP-glucosyl transferase
TraesCS5A01G394900 590540043-590541485 1.38 上调
Upregulate		 UDP-葡萄糖6-磷酸脱氢酶
UDP-glucose 6-phosphatedehydrogenase
TraesCS5A01G397600 591976101-591977663 -1.85 下调
Downregulate		 脂肪酶3类家族蛋白
Lipase class 3 family protein
TraesCS5A01G402600 595371952-595372493 -1.01 下调
Downregulate		 细胞色素6f复合亚基
Cytochrome b6f complex subunit
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