优异高粱雄性不育系01-26A的组配降秆效应及其分子机理

doi:10.3864/j.issn.0578-1752.2020.14.006

摘要/Abstract

摘要：

【目的】高粱机械化生产是未来发展的必然方向,而合理的株型是机械化生产的基础与关键。育种过程中发现,矮秆雄性不育系01-26A具有和现有粒用高粱恢复系组配F₁都能降低株高的独特优势,是一个极其难得的株型调控材料。因此,对其株高遗传效应及调控基因位点进行研究,旨在探明其株高矮化遗传机理和调控机制,以期应用遗传育种手段促进高粱株型优化提供理论依据。【方法】以具有矮化株高效应的01-26A和不具矮化株高效应的7050A高粱雄性不育系为试材,重点对其与7个（包括6个粒用和1个甜高粱）恢复系的杂种F₁的株高及节数、穗柄下茎秆高度、穗柄长和穗长等相关参数的遗传效应进行分析,同时对调控株高基因的位点Dw₁、Dw₂和Dw₃（Dw₄暂未被克隆）进行测定与分析。【结果】高粱雄性不育系01-26A（A₁细胞质）具有显著的矮化粒用高粱株高的效应,以其为母本组配的杂种F₁株高较以高粱雄性不育系7050A（A₂细胞质）为母本组配的杂种F₁株高降幅为15.8%,绝对值一般不超过160 cm,而以其为母本组配的甜高粱杂种F₁株高降低不明显,不具矮化效应;01-26A矮化株高遗传效应主要表现在杂种F₁穗柄下茎秆高度明显缩短,茎秆中下部节间长度与株高变化相关性较高;01-26A杂种F₁穗柄长降低是造成株高变矮的另一原因,其效应小于穗柄下茎秆高度,而穗长对株高变化的影响很小;通过基因位点的序列和类型分析,确定了01-26A矮化基因Dw₁—Dw₃的基因类型,并通过多个杂交组合的株高遗传数据分析,推断01-26A基因型很可能是dw₁dw₁Dw₂Dw₂dw₃dw₃dw₄dw₄,即三矮高粱不育系;另外,通过对株高调控基因研究分析,发现01-26A的dw₁和dw₃矮化基因可能对粒用高粱杂种F₁株高影响效应更大,而Dw₂的存在是造成其与甜高粱杂种F₁株高没有矮化的内在原因。【结论】高粱雄性不育系01-26A可能是具有dw₁dw₁Dw₂Dw₂dw₃dw₃dw₄dw₄基因的三矮高粱不育系,可通过降低杂种F₁穗柄下茎秆高度（主效）和穗柄长（次效）,实现高粱株高的矮化调控;但其与甜高粱杂交,可能由于Dw₂的存在,F₁并未发现明显的矮化效应。

关键词: 高粱, 株高, 遗传效应, 基因位点

Abstract:

【Objective】The mechanized production of sorghum is the inevitable direction of future development, and the ideal plant type is the basis and key factor for mechanized production. The dwarf male sterile line 01-26A was found a unique effect in reducing the plant height in F₁ generation when crossed with available grain sorghum restorer lines. Therefore, a study was conducted to determine the genetic mechanism and regulation mechanism of plant height dwarfing by its genetic effects and regulatory gene loci. 【Method】This study used sorghum male sterile line 01-26A (A₁ cytoplasm) with dwarf plant height effect and sorghum male sterile line 7050A (A₂cytoplasm) without dwarf plant height effect as the test material, focusing on 7 restorer lines, including 6 grain restorer lines and 1 sweet sorghum restorer line, and their cross F₁ generation hybrids, the genetic effects of plant height, number of nodes, the total internode length under peduncle, the peduncle length and the head length were analyzed, the gene loci of Dw₁to Dw₃ related with the plant height were also measured and analyzed. Dw₄ was not included because it had not been cloned.【Result】the male sterile line 01-26A had a significant dwarf effect on the plant height of grain sorghum, and its hybrids is 15.8% lower than that of the 7050A, generally, the absolute value of plant height did not exceed 160 cm. The plant height of F₁ generation derived from 01-26A with sweet sorghum restorer line had not been obviously reduced, so it had not dwarf effect on sweet sorghum hybrid. The genetic dwarf effect of 01-26A was mainly manifested in the shortening of the internode length under peduncle, and the internode length under peduncle has more correlated with the plant height variation. The peduncle length reduction of F₁ crossed by 01-26A with restorer lines is another reason of plant becoming shorter, but the effect was less than that of the internode length under peduncle. While the head length had much little effect on plant height variation. The dwarf genotype (Dw₁ to Dw₃) of 01-26A was determined by PCR and sequencing of Dw₁ to Dw₃ genes. And combined the analysis of plant height genetic data of multiple cross combinations, the plant height genotype of 01-26A was deduced to be dw₁dw₁Dw₂ Dw₂dw₃dw₃dw₄dw₄_, a 3-dwarf sorghum sterile line. In addition, by the analysis of plant height regulation genes, we found that the dw1 and dw3 of 01-26A may had a greater effect on the plant height of grain sorghum, while the presence of DW₂was the immanent cause for not dwarfing on sweet sorghum F₁.【Conclusion】 01-26A was likely to be a 3-dwarf sorghum male sterile line with the genotype of dw₁dw₁Dw₂Dw₂dw₃dw₃dw₄dw₄. It could achieve dwarf regulation of its F₁ by reducing the internode length (main effect) and the peduncle length (secondary effect). However, 01-26A, had not been found obvious dwarfing effect when crossed with sweet sorghum, which may be due to the presence of Dw₂.

Key words: sorghum, plant height, genetic effect, gene locus

邹剑秋,王艳秋,李金红,朱凯. 优异高粱雄性不育系01-26A的组配降秆效应及其分子机理[J]. 中国农业科学, 2020, 53(14): 2814-2827.

ZOU JianQiu,WANG YanQiu,LI JinHong,ZHU Kai. Dwarfing Effect and Molecular Mechanism of An Elite Sorghum Male Sterile Line 01-26A in Its Hybrids[J]. Scientia Agricultura Sinica, 2020, 53(14): 2814-2827.

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材料Material	系谱 Pedigree
01-26A	以7050B×P932065B为母本,以天变624B×296B×428B为父本去雄杂交,育成01-26B并经过多代回交转育而成 Take 7050B×P932065B as female parent, and Tianbian 624B×296B×428B was used as male parent to cross, the developed 01-26B was transformed into 01-26A by multiple backcross
7050A	以421B（SPL–132B）为母本,以TAM428B为父本去雄杂交,育成7050B并经过多代回交转育而成 The 421B (SPL-132B) was used as the female parent, and the TAM428B was used as the male parent to cross, the developed 7050B was transformed into by multiple backcross
LNR-4	以9544为母本,7037为父本杂交 Take 9544 as the female parent and 7037 as the male parent to cross
NK1	以LR9198为母本,0-01为父本杂交 Take LR9198 as the female parent and 0-01 as the male parent to cross
0-01	以4003×忻粱52为母本,116为父本杂交 Take 4003×Xinliang 52 as the female parent and 116 as the male parent to cross
3535	以T719为母本,0-01为父本杂交 Take T719 as the female parent and 0-01 as the male parent to cross
3550	以矮大穗为母本,LR625为父本杂交 Take short stigma as the female parent and LR625 as the male parent to cross
BR92	以F15为母本,LR9198为父本杂交 Take F15 as the female parent and LR9198 as the male parent to cross
LTR168	以辽宁地方农家品种为母本,ICSV111为父本杂交 Take Liaoning local variety as the female parent and ICSV111 as the male parent to cross

引物名称 Primer name	引物序列 Primer sequence (5′-3′)
Dw₁-F	TGGCGGTCCAACGTCTAAT
Dw₁-R	CCTGAAGTATGGCGTGTCT
Dw₂-F	CAGTTCAAATCAACGAGGAG
Dw₂-R	TCCGTCGTGAAATGAGAATA
Dw₃-F	CGTCATCGTCCAGAACTCGG
Dw₃-R	GACCCTTGCTCCACCACCTT

品种类型 Variety type	组合（母本×父本） Cross (♀×♂)	株高Plant height (cm)			F₁株高平均值 F₁ X (cm)	F₁株高变异系数 CV (%)	F₁株高与亲本关联 F₁ PH associated with the parent
品种类型 Variety type	组合（母本×父本） Cross (♀×♂)	♀	♂	平均值 Average	F₁株高平均值 F₁ X (cm)	F₁株高变异系数 CV (%)	小于低亲 PH<PH (Min)	双亲间 Between parents	大于高亲 PH> PH(Max)
	01-26A/LNR-4	103.5	145.3	124.4	141.3	13.6	—	+	—
粒用 Grain sorghum	01-26A/3550	103.5	164.8	134.2	152.5	17.4	—	+	—
	01-26A/0-01	103.5	139.2	121.4	141.9	18.7	—	—	+
	01-26A/BR92	103.5	162.4	133.0	146.6	15.3	—	+	—
	01-26A/NK1	103.5	139.1	121.3	159.7	22.9	—	—	+
	01-26A/3535	103.5	145.5	124.5	144.8	10.3	—	+	—
	均值Average	103.5	149.4	126.5	147.8	16.4
	7050A/LNR-4	136.7	145.3	141.0	173.1	18.4	—	—	+
	7050A/3550	136.7	164.8	150.8	185.7	23.7	—	—	+
	7050A/0-01	136.7	139.2	138.0	178.4	15.7	—	—	+
	7050A/BR92	136.7	162.4	149.6	179.2	16.3	—	—	+
	7050A/NK1	136.7	139.1	137.9	177.6	14.9	—	—	+
	7050A/3535	136.7	145.5	141.1	159.6	10.9	—	—	+
	均值Average	136.7	149.4	143.1	175.6	16.7
	T检验T test	*		ns	**	ns
甜高粱 Sweet sorghum	01-26A/LTR168	103.5	247.3	175.4	341.7	21.4	—	—	+
	7050A/LTR168	136.7	247.3	192.0	355.3	20.8	—	—	+
	T检验 T test	*		ns	*	ns

品种类型 Variety type	组合（母本×父本） Cross (♀×♂)	穗柄下茎秆高度 Stalk height under peduncle(cm)			F₁穗柄下茎秆高度平均值 F₁ X (cm)	F₁穗柄下茎秆高度变异系数 CV(%)	F₁穗柄下茎秆高度与亲本关联 F₁ SP associated with the parent
品种类型 Variety type	组合（母本×父本） Cross (♀×♂)	♀	♂	平均值Average	F₁穗柄下茎秆高度平均值 F₁ X (cm)	F₁穗柄下茎秆高度变异系数 CV(%)	小于低亲 SP<SP(Min)	双亲间 Between parents	大于高亲 SP> SP(Max)
	01-26A/LNR-4	47.6	82.2	64.9	75.3	14.7	—	+	—
粒用 Grain sorghum	01-26A/3550	47.6	97.4	72.5	81.6	16.9	—	+	—
	01-26A/0-01	47.6	83.3	65.5	78.1	22.1	—	+	—
	01-26A/BR92	47.6	105.6	76.6	80.6	12.9	—	+	—
	01-26A/NK1	47.6	86.9	67.3	92.7	17.4	—	—	+
	01-26A/3535	47.6	86.5	67.1	80.4	16.5	—	+	—
	均值 Average	47.6	90.3	69.0	81.5	16.8
	7050A/LNR-4	74.3	82.2	78.3	92.6	13.3	—	—	+
	7050A/3550	74.3	97.4	85.9	115.3	21.6	—	—	+
	7050A/0-01	74.3	83.3	78.8	107.7	19.5	—	—	+
	7050A/BR92	74.3	105.6	90.0	103.4	28.7	—	—	+
	7050A/NK1	74.3	86.9	80.6	101.8	23.1	—	—	+
	7050A/3535	74.3	86.5	80.4	89.7	15.4	—	—	+
	均值 Average	74.3	90.3	82.3	101.8	20.3
	T检验 T test	*		ns	**	ns
甜高粱 Sweet sorghum	01-26A/LTR168	47.6	183.4	115.5	264.6	25.2	—	—	+
	7050A/LTR168	74.3	183.4	128.9	277.2	25.6	—	—	+
	T检验 T test	*		ns	*	ns

品种类型 Variety type	组合（母本×父本） Cross(♀×♂)	穗柄长 Peduncle length (cm)			F₁穗柄长平均值 F₁ X(cm)	F₁穗柄长变异系数 CV (%)	F₁穗柄长与亲本关联 F₁ PL associated with the parent
品种类型 Variety type	组合（母本×父本） Cross(♀×♂)	♀	♂	平均值Average	F₁穗柄长平均值 F₁ X(cm)	F₁穗柄长变异系数 CV (%)	小于低亲 PL<PL(Min)	双亲间 Between parents	大于高亲 PL> PL(Max)
	01-26A/LNR-4	29.7	35.5	32.6	38.1	6.5	—	—	+
粒用 Grain sorghum	01-26A/3550	29.7	39.1	34.4	41.5	12.3	—	—	+
	01-26A/0-01	29.7	29.2	29.5	33.7	11.1	—	—	+
	01-26A/BR92	29.7	32.8	31.3	38.2	7.6	—	—	+
	01-26A/NK1	29.7	27.4	28.6	40.9	4.8	—	—	+
	01-26A/3535	29.7	31.6	30.7	36.5	8.5	—	—	+
	均值 Average	29.7	32.6	31.2	38.2	8.5
	7050A/LNR-4	33.1	35.5	34.3	49.7	6.5	—	—	+
	7050A/3550	33.1	39.1	36.1	40.3	8.7	—	—	+
	7050A/0-01	33.1	29.2	31.2	36.9	11.3	—	—	+
	7050A/BR92	33.1	32.8	33.0	46.5	7.2	—	—	+
	7050A/NK1	33.1	27.4	30.3	40.6	6.2	—	—	+
	7050A/3535	33.1	31.6	32.4	42.8	8.7	—	—	+
	均值 Average	33.1	32.6	32.9	42.8	8.1
	T检验 T test	ns		ns	*	ns
甜高粱 Sweet sorghum	01-26A/LTR168	29.7	37.3	33.5	49.9	8.1	—	—	+
	7050A/LTR168	33.1	37.3	35.2	52.1	6.5	—	—	+
	T检验 T test	ns		ns	ns	ns