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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (3): 744-754.doi: 10.3724/SP.J.1006.2023.21018

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

QTL mapping and validation of chlorophyll content of flag leaves in wheat (Triticum aestivum L.)

YANG Bin(), QIAO Ling(), ZHAO Jia-Jia, WU Bang-Bang, WEN Hong-Wei, ZHANG Shu-Wei, ZHENG Xing-Wei, ZHENG Jun()   

  1. Institute of Wheat Research, Shanxi Agricultural University / State Key Laboratory of Sustainable Dryland Agriculture (in preparation), Linfen 041000, Shanxi, China
  • Received:2022-02-25 Accepted:2022-06-07 Online:2023-03-12 Published:2022-07-07
  • Contact: ZHENG Jun E-mail:sxxmsyb83@126.com;qiaolingsmile@163.com;sxnkyzj@126.com
  • About author:First author contact:**Contributed equally to this work
  • Supported by:
    Central Guidance on Science & Technology Development of Shanxi(YDZJSX2022A033);Shanxi Agricultural University Breeding Project(YZGC013);Basic Research Program of Shanxi Province(202103021223156)

Abstract:

Flag leaf is the main photosynthetic organ in wheat. The chlorophyll content is not only the major photosynthetic pigment in flag leaf but also an important phenotypic indicator in crop breeding. Therefore, the identification of major loci/genes related to chlorophyll content in the flag leaf play an important role in breeding wheat varieties with higher grain yields and stability. In this study, we constructed a double haploid (DH) population from a cross of two cultivars with significant difference in chlorophyll content, and the chlorophyll contents of DH lines were detected under five environments. A total of 20 QTLs associated with chlorophyll content were detected using Wheat 90K single-nucleotide polymorphism (SNP) array, with contributions to phenotypic variation explained (PVE) from 4.10% to 27.16%. Three QTLs (Qchl.saw-2D.1, Qchl.saw-4D.2, and Qchl.saw-6A) were identified under multiple environmental conditions, in which Qchl.saw-2D.1 with the strongest genetic effect was different from previous studies and identified as a novel major QTL. Furthermore, Qchl.saw-2D.1 was validated by a tightly linked kompetitive allele specific PCR (KASP) marker in a recombinant inbred line (RIL) population containing the co-parent Jinmai 919. Those lines with the favorable allele of Qchl.saw-2D.1 revealed significantly higher chlorophyll content than other lines under multiple environments. Moreover, a total of 12 candidate genes controlling chlorophyll content were identified in the three QTL regions. Based on gene annotation, three genes were involved in the binding process of metal iron, such as magnesium. Five genes were regulated the structural composition of chloroplasts, and four genes were engaged in the regulation of electron transfer activities during the photosynthetic process. In conclusion, this study will broaden the understanding of the genetic mechanism and provide a molecular basis for the marker-assisted breeding (MAS) of chlorophyll content in the flag leaf of wheat.

Key words: wheat, chlorophyll content, SNP marker, QTL mapping

Table 1

Chlorophyll contents in flag leaves in parents and DHs derived from cross Jinchun 7 × Jinmai 919"

环境
Environment
亲本 Parents DH群体 DH Population
晋春7号
Jinchun 7
金麦919
Jinmai 919
平均值
Mean
标准差
SD
最大值
Max.
最小值
Min.
峰度
Kurtosis
偏度
Skewness
遗传力H2
Heritability
E1 57.07* 54.96 54.78 3.19 62.77 41.33 1.572 -0.336 0.58
E2 46.17 44.43 48.42 2.55 53.60 39.80 1.397 -0.856
E3 58.59 57.79 56.90 2.23 62.38 50.86 -0.185 -0.091
E4 55.92* 52.76 53.76 2.85 61.58 48.10 -0.392 -0.271
E5 53.59* 50.87 50.56 2.31 56.14 44.92 -0.056 -0.027
BLUP 54.57 52.38 52.88 1.62 57.23 47.64 0.258 -0.376

Fig. 1

Genetic map of DH population"

Table 2

QTLs for chlorophyll content detected in DH population"

QTL 环境
Environment
染色体
Chr.
LOD值
LOD value
表型贡献率
PVE (%)
加性效应
Additive
左标记
Left marker
右标记
Right marker
遗传位置
Genetic interval (cM)
物理位置
Physical interval (Mb)
Qchl.saw-2A.1 E4 2A 2.55 4.71 ‒0.64 Excalibur_c18630_283 Kukri_c13187_556 0.00-2.50 18.30-32.53
Qchl.saw-2A.2 E2 2A 3.33 7.10 ‒0.69 Kukri_rep_c106992_239 BS00041816_51 46.70-48.30 611.30-611.32
Qchl.saw-2A.3 E2 2A 4.98 10.33 ‒0.84 TA005399-1698 Ra_c7093_381 52.60-54.60 685.96-695.51
Qchl.saw-2A.4 E2 2A 3.04 7.75 ‒0.73 Kukri_c47259_416 RAC875_c37510_329 62.30-65.30 717.19-727.17
Qchl.saw-2B.1 E3 2B 2.95 6.97 0.59 BS00076596_51 BS00077463_51 13.30-18.20 8.34-13.76
Qchl.saw-2B.2 E2 2B 4.01 8.27 0.74 BS00105036_51 GENE-1343_556 23.60-30.20 18.18-24.10
Qchl.saw-2B.3 E4 2B 2.74 4.60 ‒0.65 Kukri_c43403_346 BS00068550_51 59.90-65.50 76.82-159.60
E5 2B 4.34 7.85 ‒0.66 Kukri_c43403_346 BS00068550_51 58.50-64.30 76.82-159.60
Qchl.saw-2D.1 E3 2D 3.69 8.21 ‒0.66 BS00012148_51 Excalibur_c18324_390 0.00-1.60 3.77-7.97
E4 2D 6.84 14.03 ‒1.11 BS00012148_51 Excalibur_c18324_390 1.10-1.60 3.77-7.97
E5 2D 12.85 26.07 ‒1.19 BS00012148_51 Excalibur_c18324_390 1.50-1.80 3.77-7.97
BLUP 2D 9.84 23.15 ‒0.78 BS00012148_51 Excalibur_c18324_390 1.40-2.10 3.77-7.97
Qchl.saw-2D.2 E1 2D 2.66 7.00 ‒1.12 BobWhite_c35859_105 GENE-1086_99 2.10-3.30 9.93-12.33
E5 2D 8.81 19.64 ‒1.22 BobWhite_c35859_105 GENE-1086_99 2.20-3.20 9.93-12.33
Qchl.saw-2D.3 E5 2D 13.60 27.16 ‒1.23 BobWhite_rep_c64429_660 D_contig04712_355 5.30-6.70 20.63-22.14
BLUP 2D 9.09 22.18 ‒0.76 BobWhite_rep_c64429_660 D_contig04712_355 5.60-6.80 20.63-22.14
Qchl.saw-2D.4 E2 2D 3.62 9.14 ‒0.78 Ex_c66554_69 D_contig73920_552 18.50-25.10 41.92-61.81
Qchl.saw-3A E3 3A 3.50 7.50 ‒0.62 BS00023222_51 Excalibur_c91430_125 123.60-129.00 714.59-724.06
E5 3A 3.12 5.30 ‒0.54 BS00023222_51 Excalibur_c91430_125 123.50-129.30 714.59-724.06
Qchl.saw-3B E4 3B 4.15 9.30 0.91 Kukri_rep_c113136_131 IACX6214 21.40-35.00 12.03-18.56
Qchl.saw-4A E2 4A 2.79 6.51 ‒0.78 Excalibur_rep_c114266_234 GENE-2399_666 15.00-15.70 612.06-618.46
Qchl.saw-4B E5 4B 3.37 5.11 ‒0.53 Kukri_c12661_326 CAP11_c1143_339 43.10-49.50 28.73-50.40
Qchl.saw-4D.1 E4 4D 2.96 5.77 0.69 Excalibur_c32965_551 Excalibur_c55561_127 22.50-24.50 43.40-119.91
BLUP 4D 2.73 4.77 0.36 Excalibur_c32965_551 Excalibur_c55561_127 13.60-24.00 43.40-119.91
Qchl.saw-4D.2 E4 4D 4.25 7.98 0.82 D_contig16493_653 CAP7_c7991_177 25.80-31.60 386.44-451.30
E5 4D 3.65 6.33 0.58 D_contig16493_653 CAP7_c7991_177 25.30-31.20 386.44-451.30
BLUP 4D 3.37 5.92 0.40 D_contig16493_653 CAP7_c7991_177 25.00-31.80 386.44-451.30
Qchl.saw-6A E2 6A 3.93 8.14 0.74 Excalibur_c1708_1975 Tdurum_contig95236_98 59.90-62.70 29.56-32.88
E4 6A 5.12 9.74 0.90 Excalibur_c1708_1975 Tdurum_contig95236_98 60.70-63.30 29.56-32.88
BLUP 6A 2.60 4.10 0.34 Excalibur_c1708_1975 Tdurum_contig95236_98 60.70-64.20 29.56-32.88
Qchl.saw-6B.1 E3 6B 2.91 6.08 ‒0.56 RAC875_c50436_101 wsnp_RFL_Contig1075_104560 79.60-100.10 686.19-694.40
Qchl.saw-6B.2 E1 6B 2.86 5.27 ‒0.74 BS00062776_51 tplb0046e21_618 116.20-116.90 706.34-708.03

Fig. 2

Additive effects of Qchl.saw-2D.1, Qchl.saw-4D.2, and Qchl.saw-6A (A) and effect of Qchl.saw-2D.1 in DH population (B)"

Table 3

Chlorophyll QTLs on chromosomes 2D, 4D, and 6A from previous studies"

染色体
Chromosome
QTL 左标记
Left marker
右标记
Right marker
物理位置
Physical position (Mb)
表型贡献率
PVE (%)
参考文献
Reference
2D qHChlb2D Xcfd53 23.03 14.31 [16]
2D QXcfd0116-2D Xcfd0116 wPt-2644 205.62 [41]
2D Qchc.iiwbr-2D Xbarc228 570.40 16.55 [42]
2D Qlchc7a.iiwbr-2D Xwmc18 130.83 31.36 [42]
2D QChl-2D Xwmc0018 wPt-0298 130.83 6.46 [40]
2D qCHN-2Da Xbcd611 Xcdo1379 4.73 [43]
2D qCHN-2Db Xfba074 Xfba062 5.49 [43]
2D QXcfd53-2D Xcfd53 23.03 [38]
2D QXbarc168.1-2D Xbarc168.1 44.74 [38]
2D QXgwm539-2D Xgwm539 513.10 [38]
4D qChlH-4D Xgwm192 Xwmc331 412.72 19.90 [39]
6A QXgwm617 Xgwm617 11.00 [22]
6A QXgwm334 Xgwm334 9.25 23.00 [22]
6A QChl-6A.1 Xgwm570 Xpsp3071 447.12 12.80 [18]
6A Qspad.acs-6A.1 Xwmc201 Xwmc684 461.84 [24]
6A Qspad.acs-6A.2 Xbarc171 Xgwm427 354.592 [24]
6A Qspad.acs-6A.3 Xgwm169 Xwmc580 595.378 [24]
6A Qspad.acs-6A.4 Xbarc113 Xwmc621 495.11 [24]
6A QChl-6A aca/caa-5 wPt-7599 32.67 7.44 [40]
6A qCHO-6A Xmwg2053 Xmwg573 7.72 [43]
6A QChl.ksu-6A Xgwm427 Xgwm169 595.38 16.10 [2]
6A QChlL.igdb‐6A Xgwm570 579.13 12.40 [44]
6A QHtscc.ksu-6A Xbarc113 CTGTGC93 17.57 [5]

Fig. 3

Genetic map of Qchl.saw-2D.1 (A), physical map of Qchl.saw-2D.1 (B), and the effect of Qchl-2D.1 in RIL population (C)"

Table 4

Screening for candidate gene information"

QTL名称 染色体 标记区间 基因 水稻同源基因 物理位置 基因注释
QTL name Chr. Marker interval Gene Rice homologous gene Position (bp) Gene annotation
Qchl.saw-2D.1 2D BS00012148_51-Excalibur_c18324_390 TraesCS2D02G007500 4038523 镁离子结合 Magnesium ion binding
Qchl.saw-2D.1 2D BS00012148_51-Excalibur_c18324_390 TraesCS2D02G008700 4685051 镁离子结合 Magnesium ion binding
Qchl.saw-2D.1 2D BS00012148_51-Excalibur_c18324_390 TraesCS2D02G010200 Os04g0118700 5230187 叶绿体RNA加工
Chloroplast RNA processing
Qchl.saw-2D.1 2D BS00012148_51-Excalibur_c18324_390 TraesCS2D02G014800 7148652 叶绿体组织 Chloroplast organization
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G273000 442898720 光系统I中的光合电子传递
Photosynthetic electron transport in photosystem I
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G248100 Os03g0212700 416933144 金属离子结合 Metal ion binding
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G270900 441653004 电子传递活性 Electron transfer activity
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G263300 Os03g0196800 434273089 叶绿体基质 Chloroplast stroma
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G277400 Os03g0163300 (OsGR1) 449725264 电子传递活性 Electron transfer activity
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G267100 Os03g0192400 437810691 电子传递链 Electron transport chain
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G230000 Os03g0240500 389572209 叶绿体外膜 Chloroplast outer membrane
Qchl.saw-4D.2 4D D_contig16493_653-CAP7_c7991_177 TraesCS4D02G275300 Os03g0161200 446137350 叶绿体 Chloroplast
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