铝毒胁迫下甘蓝型油菜种子萌发期相关性状的QTL定位

doi:10.3724/SP.J.1006.2020.94154

Abstract

Abstract:

With the aggravation of soil acidification, aluminum toxicity has become one of the important stress factors that affect crop seed germination quality and crop yield. There have been many reports on QTL mapping and candidate gene screening for aluminum tolerance related traits in crops, but few reports on QTL mapping for aluminum tolerance during germination in Brassica napus L. The seed germination test was conducted on the recombinant inbred line population treated with 80 μg mL ^-1 aluminum solution and with distilled water as control. Germination potential was investigated at 3 d, germination rate was investigated at 7 d, root length, bud length and dry weight were measured, and the relative values of various characters were calculated. Based on 6K SNP chip and combined with high density genetic linkage map, we detected 23 QTLs for five traits in rape germination period. QTLs for relative germination potential, relative germination rate, relative root length, relative bud length and relative dry weight were 9, 1, 4, 5, and 4 respectively, covering A and C genomes, with LOD values from 3.00 to 5.26, and interpretable phenotypic variation from 7.70% to 13.10%. According to the confidence interval sequence of each QTL, 30 candidate genes related to aluminum stress were screened. ALMT1 gene and MATE gene are related to the synthesis and secretion of organic acids, which mainly enhance the aluminum tolerance of plants through the secretion of organic acids such as malic acid, citric acid and oxalic acid. STOP1 gene, NAC gene and RAP2.4 gene all belong to transcription activation factors, which enhance the resistance of plants by inducing the expression of aluminum tolerance genes. ABC transporter, membrane protein transporter, GDSL lipase enhance aluminum tolerance of plants by reducing accumulation of toxic substances on plasma membrane or discharging toxic substances out of body. Peroxidase and cytochrome P450 are genes related to oxidative stress, which have the functions of preventing oxidative damage of plant cells and resisting stress. At the same time, many genes encoding stress proteins were found, which play an important role in various stress reactions. The results of this study provide a theoretical basis for the cultivation of aluminum-tolerant rape varieties and the functional research of subsequent genes.

Key words: Brassica napus L., germination, single nucleotide polymorphism, aluminum stress, QTL, candidate gene

WANG Rui-Li,WANG Liu-Yan,YE Sang,Gao Huan-Huan,LEI Wei,WU Jia-Yi,YUAN Fang,MENG Li-Jiao,TANG Zhang-Lin,LI Jia-Na,ZHOU Qing-Yuan,CUI Cui. QTL mapping of seed germination-related traits in Brassica napus L. under aluminum toxicity stress[J].Acta Agronomica Sinica, 2020, 46(6): 832-843.

Figures/Tables 7

Fig. 1

Table 1

Fig. 2

Table 2

Fig. 3

Table 3

QTL for rape germination under aluminum stress detected by composite interval mapping"

性状 Trait	QTL	标记区间 Marker interval	LOD	表观贡献率 PVE (%)	加性效应 Additive effect	置信区间 Confidence interval (cM)
相对发芽势RGV	qRGV-A01-1	AX-177912497-AX-179306831	4.33	10.90	-0.0282	37.410-43.975
	qRGV-A01-2	AX-182171878-AX-182150271	3.90	9.90	-0.0217	141.657-155.683
	qRGV-A03-1	AX-95507427-AX-177832788	4.20	10.60	0.0064	179.611-222.454
	qRGV-A03-2	AX-95504605-AX-177831059	3.74	9.50	0.0033	93.552-97.113
	qRGV-A03-3	AX-95681175-AX-95665748	3.23	8.20	-0.0236	121.496-124.157
	qRGV-A08	AX-95506362-AX-182175648	5.26	13.10	-0.0402	8.146-12.834
	qRGV-C01-1	AX-182087666-AX-95638042	3.43	8.70	-0.0123	160.226-163.199
	qRGV-C01-2	AX-177910610-AX86223402	3.00	7.70	-0.0072	1.826-6.674
	qRGV-C03	AX-95665064-AX-105309102	4.17	10.50	-0.0022	6.340-10.132
相对发芽率RGR	qRGR-C04	AX-95636072-AX-182161180	3.12	8.00	0.0329	34.378-38.45
相对根长RRL	qRRL-A03-1	AX-182087696-X-182147367	3.14	8.00	-0.0349	144.851-146.986
	qRRL-A03-2	AX-95509347-AX-95509401	3.11	8.00	-0.0366	207.579-211.5
	qRRL-A09	AX-95635531-AX-182177329	3.36	8.60	-0.0348	59.898-79.773
	qRRL-C03	AX-95662770-AX-177910217	3.23	8.20	-0.0274	218.487-221.508
相对芽长RBL	qRBL-A03	AX-95636441-AX-177911244	3.12	8.00	-0.0038	32.513-47.814
	qRBL-A08-1	AX-95663434-AX-95506362	3.34	8.50	-0.0158	2.275-8.146
	qRBL-A09-1	AX-177912380-AX-177829538	4.07	10.30	0.0216	178.878-188.216
	qRBL-A09-2	AX-177832428-AX-182127697	3.53	9.00	0.0224	6.106-15.138
	qRBL-C08	AX-95665611-AX-86227922	4.75	11.90	0.0115	94.661-132.074
相对干重RDW	qRDW-A09-1	AX-182158057-AX-182154686	3.60	7.80	0.0196	127.938-128.931
	qRDW-A09-2	AX-95682972-AX-182133711	3.05	9.10	0.0067	275.002-280.965
	qRDW-A10-1	AX-182087443-AX-65636575	4.09	10.30	0.0050	129.25-131.783
	qRDW-A10-2	AX-177829985-AX-182146546	3.90	9.90	0.0036	160.157-160.711

Table 3

Table 4

Comparison between QTL confidence interval candidate genes in Brassica napus genome and Arabidopsis stress related genes"

性状 Trait	物理区间 Physical interval	甘蓝型油菜基因编号 Gene ID in B. napus	基因登录号 Gene accession	基因注释 Gene annotation	参考文献 Reference
相对发芽势RGV	18835423-22795371	BnaA01g26740D	AT3G18440	Aluminum-activated malate transporter 9 (ALMT9)	[14]
	20061150-26605700	BnaA03g57410D	AT3G06130	Heavy metal-associated domain (HMA)	[23]
		BnaA03g39800D	AT5G61240	Leucine-rich repeat (LRR) family protein	[24]
	14408144-15038093	BnaA03g03270D	AT5G11250	Disease resistance protein (TIR-NBS-LRR class)
		BnaA03g14650D	AT2G31660	Super sensitive to ABA and drought2 (SAD2)	[25]
	11435749-11630658	BnaA03g13730D	AT2G30140	UDP-Glycosyltransferase superfamily protein	[26]
	1012939-2022032	BnaA08g30510D	AT4G19960	K⁺ uptake permease 9 (KUP9)	[27]
		BnaA08g19190D	AT1G24620	EF hand calcium-binding protein family (CML25)	[28]
		BnaA08g26970D	AT1G08430	Aluminum-activated malate transporter 1 (ALMT1)
	676816-1129401	BnaC01g23380D	AT3G51490	Metallothionein 3 (MT3)	[29]
		BnaC01g01650D	AT4G37270	Response to toxic substance (HMA)
	3925832-4284222	BnaC03g08920D	AT5G18370	Disease resistance protein (TIR-NBS-LRR class) family	[30]
相对发芽率RGR	3627529-9882343	BnaC04g12580D	AT2G27780	Transcription factor IIS family protein	[31]
相对根长RRL	17577932-2005963	BnaA03g35920D	AT3G20750	Zinc ion binding (GATA)	[32]
	24406111-29734315	BnaA03g35900D	AT5G26717	Putative membrane lipoprotein	[33]
		BnaA03g54580D	AT1G34370	Sensitive to proton rhizotoxicity 1 (STOP1)	[15]
	866809-2227131	BnaA09g11920D	AT1G64065	Late embryogenesis abundant (LEA)	[34]
		BnaA09g14610D	AT3G43570	GDSL-like Lipase/Acylhydrolase superfamily protein	[35]
	51210223-55837858	BnaC03g04310D	AT5G44050	MATE efflux family protein	[16]
		BnaC03g65810D	AT4G34710	Response to osmotic stress (ADC2)	[36]
相对芽长RBL	3249677-4604524	BnaA03g39360D	AT5G02490	Heat shock protein 70 (Hsp 70) family protein	[37]
	598224-1012939	BnaA08g01480D	AT1G52490	F-box associated interaction domain	[38]
	22081699-23394648	BnaA09g02290D	AT3G28415	ABC transporter family protein	[39]
		BnaA09g30790D	AT1G22220	F-box family protein
		BnaA09g30810D	AT1G22190	Response to toxic substance (RAP2.4)	[40]
	575326-705822	BnaA08g28220D	AT1G05250	Peroxidase superfamily protein	[41]
	19580994-23049951	BnaC08g07240D	AT1G33110	MATE efflux family protein
相对干重RDW	32408555-33029777	BnaA09g48330D	AT1G49360	F-box family protein
	16531371-16654826	BnaA10g24620D	AT3G26180	Cytochrome P450 proteins	[42]
		BnaA10g25760D	AT5G04410	Regulation of flavonoid biosynthetic process (NAC)	[43]

Table 4

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相关性状 Relative trait	亲本 Parents		RIL群体 RIL population
相关性状 Relative trait	ZS11	10D130	均值Mean	极差Range	标准差SD	变异系CV(%)
相对发芽势 RGV	0.982	0.967	1.032^**	1.667	0.230	22.3
相对发芽率 RGR	0.983	0.982	1.043^**	1.030	0.140	13.4
相对根长 RRL	0.621	0.472	0.465^**	0.733	0.146	31.3
相对芽长 RBL	0.955	0.987	0.994^**	1.158	0.178	17.9
相对干重 RDW	0.987	0.990	0.920^**	1.125	0.175	19.0

QTL mapping of seed germination-related traits in Brassica napus L. under aluminum toxicity stress

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测定指标Index	相对发芽势RGV	相对发芽率RGR	相对根长RRL	相对芽长RBL	相对干重RDW
相对发芽势RGV	1
相对发芽率RGR	0.404^**	1
相对根长RRL	0.101	0.053	1
相对芽长RBL	-0.108	-0.109	0.062	1
相对干重RDW	-0.148^*	-0.252^**	-0.075	-0.020	1

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