小麦旗叶叶绿素含量的QTL定位及验证

doi:10.3724/SP.J.1006.2023.21018

Abstract

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

YANG Bin, QIAO Ling, ZHAO Jia-Jia, WU Bang-Bang, WEN Hong-Wei, ZHANG Shu-Wei, ZHENG Xing-Wei, ZHENG Jun. QTL mapping and validation of chlorophyll content of flag leaves in wheat (Triticum aestivum L.)[J].Acta Agronomica Sinica, 2023, 49(3): 744-754.

Figures/Tables 7

Table 1

Fig. 1

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

Table 2

Fig. 2

Table 3

Fig. 3

Table 4

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环境 Environment	亲本 Parents		DH群体 DH Population
环境 Environment	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

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]

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

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

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