甘蓝型春油菜早花位点加密及位点聚合创建优异早花资源

doi:10.3724/SP.J.1006.2020.04012

作物学报 ›› 2020, Vol. 46 ›› Issue (11): 1711-1721.doi: 10.3724/SP.J.1006.2020.04012

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

甘蓝型春油菜早花位点加密及位点聚合创建优异早花资源

柳海东(), 潘云龙(), 杜德志^*()

青海大学农林科学院 / 青海省春油菜遗传改良重点实验室 / 国家油菜改良中心青海分中心 / 农业农村部春油菜科学观测实验站 / 青海省春油菜工程技术研究中心, 青海西宁 810016

收稿日期:2020-01-15 接受日期:2020-06-02 出版日期:2020-11-12 网络出版日期:2020-06-15
通讯作者: 杜德志
作者简介:柳海东, E-mail: dahaima@163.com|潘云龙, E-mail:pylscorpio@126.com
基金资助:
本研究由国家重点研发计划项目(2018YFD0100501);国家自然科学基金项目(31760395);国家现代农业产业技术体系建设专项(CARS-12);青海省重点研发与转化计划项目(2018-NK-C07);青海省春油菜遗传改良重点实验室(2017-ZJ-Y09);青海省农林科学院科研专项(2018-NKY-011)

Locus encryption for early flowering and QTL polymerization to create excellent early flowering resources of spring Brassica napus L.

LIU Hai-Dong(), PAN Yun-Long(), DU De-Zhi^*()

Academy of Agricultural and Forestry Sciences, Qinghai University / Qinghai Spring Rape Engineering Research Center / Spring Rape Scientific Observation Experimental Station of Ministry of Agriculture and Rural Areas / Qinghai Research Branch of the National Rapeseed Genetic Improvement Center / Key Laboratory of Qinghai Province for Spring Rapeseed Genetic Improvement, Xining 810016, Qinghai, China

Received:2020-01-15 Accepted:2020-06-02 Published:2020-11-12 Published online:2020-06-15
Contact: De-Zhi DU
Supported by:
This study was supported by the National Key Research and Development Plan(2018YFD0100501);the National Natural Science Foundation of China(31760395);the China Agriculture Research System(CARS-12);the Key Research and Development and Transformation Project of Qinghai Province(2018-NK-C07);the Laboratory of Spring Rape Genetic Improvement of Qinghai Province(2017-ZJ-Y09);and the Qinghai Academy of Agriculture and Forestry Sciences Research Project(2018-NKY-011)

摘要/Abstract

摘要：

前期在甘蓝型春油菜NNDH群体中定位到cqDTFA7a和cqDTFC8 2个早花主效QTL, 分别开发了与cqDTFA7a紧密连锁的SSR标记G1803、InDel标记IA7-4, 以及与cqDTFC8紧密连锁的SSR标记S035。本研究在此基础上构建了早花QTL位点cqDTFC8的BC₂F₂群体, 进一步对该位点加密, 开发了与cqDTFC8紧密连锁的标记SNP11。利用开发的与两个位点紧密连锁的4个标记对93个甘蓝型春油菜自然资源进行早花基因型鉴定, 从中筛选出含有cqDTFA7a位点的资源3164和2216, 含有cqDTFC8位点的资源3484和2857, 两位点资源之间两两正反杂交进行位点聚合。通过小孢子培养和标记辅助选择快速获得聚合DH系, 筛选出性状优良且早花的聚合系与波里马细胞质雄性不育系配置杂交组合, 连续2年多点进行测产试验, 对聚合系利用价值进一步分析。cqDTFC8加密结果显示, 该位点被定位于SNP11和SNP12区间中, 与SNP11共分离。自然资源早花基因型鉴定结果显示, 含有cqDTFA7a位点的单株50个, 平均初花期58.1 d, 含有cqDTFC8位点的单株16个, 平均花期58.3 d, 同时含有两位点的单株16个, 平均花期55.2 d, 表明同时包含两早花位点株系比只含有一个早花位点株系早开花。聚合结果显示, 早花位点cqDTFC8和cqDTFA7a的聚合单株开花时间比单位点亲本单株的开花时间提前2~3 d, 其中来自cqDTFA7a位点的3164和cqDTFC8位点的3484聚合后的DH18比亲本早开花3 d, 产量相关性状都优于其他品系。进一步利用DH18与波里马细胞质雄性不育系025A配置组合, 将该组合定名为TZG18, 2年9点测产结果表明, TZG18比青藏高原白菜型主栽品种浩油11号增产17.5%以上。综上研究结果说明, 早花位点聚合品系比早花单位点品系在开花时间上有着明显的优势, 同时对增加油菜产量也有影响。本研究是甘蓝型春油菜早花性状MAS育种的初步探索, 为春油菜区特早熟甘蓝型春油菜品种替代白菜型油菜提供了材料支持, 也为基因聚合育种技术提供了新途径。

关键词: 甘蓝型春油菜, 早花位点, 基因聚合, 早花资源

Abstract:

cqDTFA7a and cqDTFC8, two major effects of early flowering QTL, were identified in the NNDH population of spring Brassica napus, and closely linked markers SSR G1803, InDel IA7-4, and SSR S035 with cqDTFC8 developed in previous studies. In this study, a BC₂F₂ population for early flowering QTL locus cqDTFC8 was constructed, and a closely linked SNP marker was further developed. The early flowering genotypes of one natural resources contained 93 spring B. napus varieties were identified used four closely linked markers with two loci, and selected 3164 and 2216 resources with cqDTFA7a site, 3484 and 2857 resources with cqDTFC8 site. Two site resources were aggregated by site polymerization through reciprocal hybridization. The polymerized DH system was rapidly obtained by microspore culture and maker assisted selection. A hybrid combination was created between polymeric line with good traits and early flowers and the Polima CMS, and the utilization value of the polymeric line was further analyzed by the production test at multiple environments for two consecutive years. cqDTFC8 encryption results showed that this site was located in the SNP11 and SNP12 interval, and separated from SNP11 altogether. The identification results of early flowering genotypes of natural resources showed that there were 50 individuals containing cqDTFA7a locus, with an average initial flowering period of 58.1 days; 16 single plants containing cqDTFC8 locus, with an average flowering period of 58.3 days; and 16 single plants containing two loci, with an average flowering period of 55.2 days, indicating the more early flowering sites containing, the earlier flowering. The results of polymerization showed that the flowering time of the polymerized lines of cqDTFC8 and cqDTFA7a was 2-3 days earlier than that of the single locus parents, among which the polymerized DH18 from 3164 of cqDTFA7a and 3484 of cqDTFC8 was 3 days earlier than that of the parents, and the yield-related traits were better than those of other lines. The combination of DH18 and the Polima CMS 025A was further utilized, and the combination was named TZG18. Yield results of two years and nine environments showed that yield of TZG18 was above 17.5% higher than the Haoyou 11, a local B. rapa variety on the Qinghai-Tibet plateau. Those results indicated that the early flowering site polymeric lines had an obvious advantage over the single locus lines in flowering time, and had an effect on the increasement of rapeseed yield. This study is a preliminary exploration of MAS breeding for early flowering traits of Brassica napus, providing materials support for replacing B.rapa varieties using early maturity Brassica napus varieties in spring rapeseed region, and approaches for gene polymerization breeding technology.

Key words: spring Brassica napus, early flowering locus, gene pyramiding, early flowering resources

柳海东, 潘云龙, 杜德志. 甘蓝型春油菜早花位点加密及位点聚合创建优异早花资源[J]. 作物学报, 2020, 46(11): 1711-1721.

LIU Hai-Dong, PAN Yun-Long, DU De-Zhi. Locus encryption for early flowering and QTL polymerization to create excellent early flowering resources of spring Brassica napus L.[J]. Acta Agronomica Sinica, 2020, 46(11): 1711-1721.

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引物 Primer	位置 Position (bp)	突变、插入/缺失 Mutate, insert/miss	上游引物序列 Forward primer (5°-3°)	下游引物序列 Reverse primer (5°-3°)
SNP6	1636463	G/A	CCAGACCCAACATTACCC	TCCGGTGTTCTGCTTATT
SNP11	2777872	G/A	GAAAGCTGCTGGGTTTCT	TATTATGCCATGTCATCTAC
SNP12	2833887	A/G	CTCGGACGAACAGAACCC	TCAGCTCATGGAGGAACA
SNP14	3147552	T/C	TTGAAGTCGTGCGTCGGT	AGCAGAGTAGAGGAAGCTGGTA
SNP20	3674261	T/C	GAGACAAATCTCAGCAGT	CAAGATGACGAATCCAAT
SNP21	3706251	T/C	GTTTATGTCTGACCTTGTT	GTGGAACTTGATGGATTT
InDel 53	5405790	CA/C	ATAGACGTAGAGGCTGAGCA	AATCTCGTCACCATCAAGAG
InDel 55	5409462	GAATAACATAAAACTT/G	GGTGTTCATCTCTGAAGACAA	TTGTGATTTGTGAATTGATGA
InDel 60	5527563	GTCGC/G	CATGCTTCATCTCTTCAACA	GTCAACCAGCGAGTAGAAAC

基因型Genotype				资源数 Number of resources	初花期First flowering period (d)
cqDTFA7a		cqDTFC8		资源数 Number of resources	变幅 Range	平均值±标准差 Mean±SD	P值 P-value
G1803	IA7-4	S035	SNP11
AA				56	53.2-62.1	58.7±4.3	0.04^a
BB				37	63.2-77.3	65.7±5.2
	AA			50	54.2-62.3	58.1±3.9	0.04^b
	BB			43	63.5-78.2	65.9±4.8
		AA		21	55.7-61.7	58.6±3.6	0.04^c
		BB		72	62.3-77.7	64.9±4.1
			AA	16	52.4-59.3	58.3±3.7	0.04^d
			BB	77	62.1-79.3	64.4±5.1
AA	AA	AA	AA	16	53.2-57.6	55.2±3.2	0.008^e
BB	BB	BB	BB	30	65.4-79.3	65.0±4.6

材料名称 Material name	位点 Locus	来源 Source	农艺性状Agronomic trait					开花时间 Flowering time (d)
材料名称 Material name	位点 Locus	来源 Source	角果长 Fruit length (cm)	每角粒数 Seed number	千粒重 Thousand seeds weight (g)	全株角果数 Carob number	单株产量 Yield per plant (g)	开花时间 Flowering time (d)
3164	cqDTFA7a		7.8±1.9 bc	29.6±1.0 a	3.8±0.2 cd	293±28.0 bc	16.7±2.4 b	58.3±2.2 abc
2216	cqDTFA7a		7.6±1.2 c	25.0±2.5 bcd	4.2±0.1 bcd	274±40.5 cd	16.5±3.4 b	59.2±2.1 ab
3484	cqDTFC8		8.0±1.0 ab	27.2±3.0 b	4.2±0.3 abc	341±10.1 ab	17.5±3.7 ab	57.6±2.3 abc
2857	cqDTFC8		6.9±1.1 e	26.3±3.0 bc	3.6±0.2 f	271±14.5 cde	14.7±2.6 b	60.0±3.0 a
DH18	cqDTFC8 & cqDTFA7a	3484×3164	8.1±1.1 a	32.0±2.0 a	4.5±0.1 a	354±26.6 a	21.4±2.1 a	54.5±1.2 c
DH2	cqDTFA7a & cqDTFC8	2216×3484	7.0±1.9 e	23.1±5.0 d	4.1±0.1 cd	264±29.5 cde	18.5±2.2 ab	55.6±2.0 bc
DH20	cqDTFA7a & cqDTFC8	3164×3484	7.2±1.1 d	23.6±7.0 cd	3.6±0.2 ef	339±37.0 ab	17.1±2.0 ab	55.8±1.2 bc
DH12	cqDTFA7a & cqDTFC8	2216×2857	7.1±1.6 de	23.4±3.0 cd	4.3±0.1 abc	257±38.5 sdef	16.3±3.2 b	56.2±2.1 abc
DH5	cqDTFA7a & cqDTFC8	3164×2857	6.8±0.9 e	27.6±3.5 b	3.9±0.4 de	216±21.3 f	16.4±2.2 b	56.4±1.2 abc
DH4	cqDTFC8 & cqDTFA7a	3484×2216	6.5±0.4 e	25.2±5.1 bcd	4.1±0.1 bcd	239±17.2 def	15.8±3.1 b	56.8±1.4 abc
DH23	cqDTFC8 & cqDTFA7a	2857×3164	6.8±0.1 e	27.8±3.2 b	4.4±0.2 ab	225±22.5 ef	16.7±4.1 b	57.3±1.3 abc

品种 Variety	产量Yield (kg hm^-2)						平均 Average	比对照增产 Yield increase over control (%)
品种 Variety	门源沙沟梁 Menyuan shagouliang	门源后院 Menyuan backyard	湟源 Huangyuan	同德 Tongde	贵南牧场 Guinan Ranch	海北 Haibei	平均 Average	比对照增产 Yield increase over control (%)
2018
TZG18	620.0±80.0 a	980.0±166.7 a	2233.3±100.0 a	1733.3±133.3 a			1391.7±73.3 a	21.0
浩油11号 Haoyou 11 (CK)	686.7±93.3 a	1113.3±160.0 a	1986.7±140.0 b	813.3±60.0 b			1150.0±53.3 b	0
2019
TZG18		1746.7±160.0 a	2960.0±206.7 a	2500.0±106.7 a	2453.3±140.0 a	2780.0±233.3 a	2488.0±113.3 a	17.5
浩油11号 Haoyou 11 (CK)		1146.7±0.14 b	2393.3±186.7 b	2213.3±80.0 b	2566.7±153.3 a	2266.7±160.0 b	2117.3±93.3 b	0

甘蓝型春油菜早花位点加密及位点聚合创建优异早花资源

Locus encryption for early flowering and QTL polymerization to create excellent early flowering resources of spring Brassica napus L.

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