油菜高油酸种质的创建及高油酸性状遗传与生理特性的分析

doi:10.3864/j.issn.0578-1752.2021.02.003

摘要/Abstract

摘要：

【目的】创建高油酸（high oleic,HO）油菜新种质,探明HO新种质中高油酸性状的遗传模式,明确HO新种质油酸含量变化的生理特性,为培育HO油菜新品种奠定基础。【方法】采用辐射处理油菜萌动发芽种子,获得初级诱变群体后在后续世代利用极端选择法结合小孢子培养技术筛选油菜HO新种质。以HO新种质分别与3个不同遗传背景的常规油菜品系为亲本组合杂交构建6个世代（P₁、P₂、F₁、BC₁P₁、BC₁P₂和F₂）的遗传群体,测定各群体脂肪酸含量后应用主基因+多基因混合遗传模型联合分析方法对遗传群体的高油酸性状进行遗传分析。测定HO新种质种子发芽过程中子叶、不同温度处理下苗期营养器官以及角果成熟过程中种子的油酸含量,明确其变化规律及生理效应。【结果】通过辐射获得油酸含量显著变化的初级诱变群体,在后续世代持续利用极端选择法得到平均油酸含量升高20个百分点的高世代群体,采用小孢子培养得到纯合稳定的油菜HO双单倍体群体,最终根据品质性状筛选成功得到HO新种质B161,其油脂中油酸含量为85%,亚麻酸含量为3%。以B161为HO亲本和常规品系杂交配置得到3个具有不同遗传背景的遗传群体并测定获得各群体的油酸含量表型数据。脂肪酸含量相关性分析表明,十八碳脂肪酸中油酸含量与亚油酸含量和亚麻酸含量具有显著负相关关系。遗传分析结果表明,该种质中高油酸性状由2对具有加性效应的主效基因控制,并且2对基因对油酸含量的效应值接近。生理分析表明,常温下HO品系营养器官（根、茎、叶和叶柄）的油酸含量均显著高于常规品系,亚麻酸含量显著低于常规品系。低温下HO品系营养器官的油酸含量降低,但仍高于常规品系;常规品系油酸含量在低温下稳定。低温下两类品系营养器官的亚麻酸含量均显著提高,但HO品系亚麻酸含量仍低于常规品系。在种子成熟过程和种子发芽过程中,HOLL品系中油酸含量持续显著高于常规油菜,而亚麻酸含量则持续显著低于常规油菜。【结论】成功创制了油菜HO新品系B161,明确了新种质HO性状的遗传规律和生理特性。获得的HO品系具有潜在育种利用价值。

关键词: 油菜, 辐射诱变, 种质创新, 高油酸性状, 遗传分析, 低温响应

Abstract:

【Objective】 This study is to create the new high oleic (HO) canola germplasm, to explore its genetic mode and the physiological characters of the HO trait, which will lay a foundation for breeding HO canola varieties. 【Method】 The primary mutation population with was obtained by radiation treatment of germinating canola seeds. The new HO germplasm was screened by extreme selection method combined with microspore culture technology in subsequent generations. The genetic populations of six generations (P₁, P₂, F₁, BC₁P₁, BC₁P₂ and F₂) were constructed by crossing the HO Germplasm with three conventional canola lines with different genetic background. After the fatty acid content of each population was determined, the genetic analysis of high oleic acid content in the genetic population was analyzed by the mixed major-gene plus polygene inheritance model. The oleic acid content in cotyledons during seed germination, vegetative organs at seedling stage in different temperature regimes and seeds during silique ripening process of the HO germplasm were detected to explore their change patterns and physiological effects.【Result】 The primary mutation population with significantly increased oleic acid content was obtained by radiation treatment, and then the high generation population with 20-percent-increased oleic acid content was obtained by using extreme selection method in subsequent generations and the double haploid population was obtained by microspore culture. Finally, a new HO germplasm B161 (C18:1=85%, C18:3=3%) was successfully screened according to the quality traits. Three genetic populations with different genetic background were obtained by crossing B161 as HO parent with three other conventional lines. The correlation analysis of fatty acid contents showed that there was a significant negative correlation between oleic acid content, linoleic acid content and linolenic acid content. The results of genetic analysis showed that the high oleic acid content was controlled by two major genes with additive effect, and their effect values were close. Physiological analysis showed that the contents of oleic acid in vegetative organs (root, stem, leaf and petiole) of HO line were significantly higher than those of the conventional strain, and the linolenic acid contents of HO line were significantly lower than those of the conventional line. The contents of oleic acid in vegetative organs of HO line decreased at low temperature, but they were still higher than those of the conventional line. The linolenic acid contents in vegetative organs of the two lines increased significantly at low temperature, but the linolenic acid content of HO line was still lower than that of the conventional line. During the process of seed ripening and seed germination, the oleic acid content of HO line was significantly higher than that of conventional line, while the linolenic acid content was significantly lower than that of conventional line.【Conclusion】The new HO germplasm was successfully created and the genetic mode and physiological characters were confirmed. This HO germplasm has the potential value in breeding.

Key words: Brassica napus L., radiation mutagenesis, germplasm creation, high oleic trait, genetic analysis, response to low temperature

龙卫华,浦惠明,高建芹,胡茂龙,张洁夫,陈松. 油菜高油酸种质的创建及高油酸性状遗传与生理特性的分析[J]. 中国农业科学, 2021, 54(2): 261-270.

LONG WeiHua,PU HuiMing,GAO JianQin,HU MaoLong,ZHANG JieFu,CHEN Song. Creation of High-Oleic (HO) Canola Germplasm and the Genetic and Physiological Analysis on HO Trait[J]. Scientia Agricultura Sinica, 2021, 54(2): 261-270.

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https://www.chinaagrisci.com/CN/Y2021/V54/I2/261

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年度 Year	世代 Generation	样本数 No. of sample	C18:1			C18:3
年度 Year	世代 Generation	样本数 No. of sample	平均值 Mean	变幅 Range (%)	变异系数 CV (%)	平均值 Mean	变幅 Range (%)	变异系数 CV (%)
2004	M₁	201	73.44	68.75—78.81	2.19	5.38	3.20—7.91	20.24
2005	M₂	578	73.97	56.14—84.90	4.50	6.02	3.52—8.97	32.63
2006	M₃	248	75.44	65.50—85.99	5.42	4.30	1.87—8.48	36.91
2007	M₄	217	78.76	69.36—87.19	9.69	3.92	1.57—14.02	45.04
2008	M₅	208	83.30	70.54—86.95	4.99	3.10	1.98—7.95	38.68
2009	M₆	65	81.23	73.51—85.47	2.36	4.00	2.54—6.38	14.20

世代 Generation	籽粒数 No. of seeds	脂肪酸 Fatty acid
世代 Generation	籽粒数 No. of seeds	C16:0	C18:0	C18:1	C18:2	C18:3	C20:1	C22:1
P₁	16	3.10±0.31	2.10±0.32	85.36±0.50	5.03±0.21	2.52±0.11	1.25±0.13	0.20±0.05
P₂	17	3.61±0.35	1.59±0.13	64.53±0.72	21.59±0.59	7.94±0.21	1.22±0.14	0.21±0.03
F₁(P₁×P₂)	38	3.28±0.29	1.72±0.22	73.98±1.29	11.66±0.90	4.93±0.42	1.28±0.17	0.19±0.05
F₁(P₂×P₁)	40	3.43±0.31	1.69±0.19	74.14±1.56	11.41±1.30	4.94±0.54	1.31±0.18	0.20±0.04
BC₁P₁	95	3.36±0.39	1.86±0.20	79.57±4.52	9.30±4.07	3.89±1.01	1.33±0.16	0.29±0.16
BC₁P₂	98	3.40±0.35	1.53±0.18	70.84±3.61	15.11±3.40	6.90±1.24	1.32±0.11	0.27±0.15
F₂	198	3.21±0.43	1.63±0.24	74.95±5.20	12.68±4.48	5.34±1.37	1.38±0.19	0.25±0.20

脂肪酸 Fatty acid	棕榈酸C16:0	硬脂酸C18:0	油酸C18:1	亚油酸C18:2	亚麻酸C18:3	二十碳烯酸C20:1	芥酸C22:1
C16:0	1	-0.106	-0.474**	0.409**	0.222**	-0.371**	0.029
C18:0		1	0.029	-0.037	-0.143*	0.033	-0.010
C18:1			1	-0.943**	-0.541**	0.204**	-0.062
C18:2				1	0.303**	-0.330**	-0.007
C18:3					1	-0.092	-0.005
C20:1						1	0.178*
C22:1							1

组合 Combination	C18:1
组合 Combination	备选模型 Model	极大似然值 Max-likelihood-value	AIC值 AIC value
B161×N137	2MG-ADI	-597.9275	1215.855
	2MG-EA	-607.3177	1220.635
	1MG-A	-607.3178	1220.636
B161×N27	2MG-A	-586.1464	1180.293
	2MG-ADI	-582.3989	1184.798
	1MG-AD	-590.8457	1189.691
B161×N15	2MG-A	-576.5989	1161.198
	1MG-AD	-582.3780	1172.756
	2MG-EAD	-583.5032	1173.006

一阶参数 Univalent parameter	估计值 Estimate value
一阶参数 Univalent parameter	B161×N137	B161×N27	B161×N15
第1对主基因的加性效应da	5.6587	4.7107	4.3096
第2对主基因的加性效应db	4.7263	6.3145	4.2901
第1对主基因的显性效应ha	/	/	/
第2对主基因的显性效应hb	/	/	/