Scientia Agricultura Sinica ›› 2020, Vol. 53 ›› Issue (14): 2814-2827.doi: 10.3864/j.issn.0578-1752.2020.14.006

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• SPECIAL FOCUS: SORGHUM BREEDING AND CULTIVATION • Previous Articles     Next Articles

Dwarfing Effect and Molecular Mechanism of An Elite Sorghum Male Sterile Line 01-26A in Its Hybrids

ZOU JianQiu(),WANG YanQiu,LI JinHong,ZHU Kai()   

  1. Sorghum Research Institute, Liaoning Academy of Agricultural Sciences, Shenyang 110161
  • Received:2019-07-31 Accepted:2019-09-22 Online:2020-07-16 Published:2020-08-10
  • Contact: Kai ZHU E-mail:jianqiuzou@126.com;zhukai72@163.com

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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 (A2 cytoplasm) 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 Dw1 to 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 DW2 was 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

Table 1

The pedigree of two sorghum male sterile lines and seven sorghum restorer lines"

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

Table 2

PCR primer amplification sequence"

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

Table 3

Genetic effects of plant height of sterile lines, restorer lines and F1 hybrids"

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
♀ ♂ 平均值
Average		 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

Table 4

Genetic effects of stalk height under the peduncle of male sterile lines, restorer lines and F1 hybrids"

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
♀ ♂ 平均值Average 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

Table 5

Genetic effects of peduncle length of male sterile lines, restorer lines and F1 hybrids"

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
♀ ♂ 平均值Average 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

Table 6

Genetic effects of head length of male sterile lines, restorer lines and F1 hybrids"

品种类型
Variety type		 组合（母本×父本）
Cross (♀×♂)		 穗长 Head length (cm) F1代穗长
平均值
F1 X (cm)		 F1代穗长
变异系数
CV (%)		 F1穗长与亲本关联 F1 HL associated with the parent
♀ ♂ 平均值Average 小于低亲
HL<HL(Min)		 双亲间
Between parents		 大于高亲
HL> HL(Max)
01-26A/LNR-4 26.2 27.6 26.9 27.9 2.1 — — +
粒用
Grain sorghum		 01-26A/3550 26.2 28.3 27.3 29.4 1.4 — + —
01-26A/0-01 26.2 26.7 26.4 30.1 4.5 — — +
01-26A/BR92 26.2 24.0 25.1 27.8 5.3 — — +
01-26A/NK1 26.2 24.8 25.4 26.1 2.9 — — +
01-26A/3535 26.2 27.4 26.7 27.9 3.2 — — +
均值 Average 26.2 26.5 26.3 28.2 3.2
7050A/LNR-4 29.3 27.6 28.5 30.8 4.2 — — +
7050A/3550 29.3 28.3 28.8 30.1 2.6 + — —
7050A/0-01 29.3 26.7 28.0 33.8 3.9 — + —
7050A/BR92 29.3 24.0 26.7 29.3 7.8 — — +
7050A/NK1 29.3 24.8 27.1 35.2 5.2 — + —
7050A/3535 29.3 27.4 28.4 27.1 3.1 + — —
均值 Average 29.3 26.5 27.9 31.1 4.5 — + —
T检验 T test ns ns ns *
甜高粱Sweet sorghum 01-26A/LTR168 26.2 26.6 26.4 27.2 3.7 + — —
7050A/LTR168 29.3 26.6 27.9 26.0 2.9 + — —
T检验 T test ns ns ns *

Table 7

Genetic effect of relevant parameters of stalk to plant height of F1 hybrids"

品种类型
Variety type		 组合（母本×父本）
Cross (♀×♂)		 与株高相关系数R Correlation coefficient with plant height R (%)
节间长Internode length 节数
Internode number
下层Lower 中下层Lower middle 中层Middle 中上层Up middle 上层Upper
粒用
Grain sorghum		 01-26A/LNR-4 15.3 65.2* 52.6* 18.7 22.3 75.3*
01-26A/3550 -3.6 42.6 63.2* 55.6* 45.2* 64.2*
01-26A/0-01 34.5 51.8* 71.8** 23.4 12.7 26.9
01-26A/BR92 -12.4 63.2* 51.4* 42.3* 26.9 65.8*
01-26A/NK1 12.3 68.2* 76.3** 9.8 23.7 69.7*
01-26A/3535 23.5 42.3 55.2* 48.5 14.3 54.8*
7050A/LNR-4 5.3 29.7 38.5 68.9* 24.1 76.7*
7050A/3550 -2.9 15.4 25.6 45.6* 82.5** 72.3*
7050A/0-01 14.3 42.4 42.1 68.7** 43.6 71.9*
7050A/BR92 21.6 28.5 23.6 76.3* 67.1* 52.3*
7050A/NK1 12.3 9.7 -9.7 77.9** 69.9** 88.9**
7050A/3535 18.5 41.2 45.3 56.2* 48.7 65.3*
甜高粱
Sweet sorghum		 01-26A/LTR168 8.4 31.4 55.9* 75.3** 42.1 61.5*
7050A/LTR168 9.7 15.6 58.4* 65.3* 28.6 65.3*

Fig. 1

Verification of the dwarf effect of F1 hybrids"

Fig. 2

Analysis of Dw1 in sorghum"

Fig. 3

Analysis of Dw2 gene locus in sorghum **: Gene deletion; ··: Heterozygosity"

Fig. 4

Analysis of Dw3 in sorghum 1, 2, 3, 7, 9, 10, 12, 13, 14, 20, 21 and 23 are 1263 bp in size and the other fragments are 2145 bp in size"

Table 8

Dw3 test serial number and its combination"

序号
Serial number		 品系或组合
Line or combination		 基因型
Genotype		 序号
Serial number		 品系或组合
Line or combination		 基因型
Genotype
1 LNR-4 Dw3Dw3 13 7050A/LNR-4 Dw3dw3
2 3550 Dw3Dw3 14 7050A/BR92 Dw3dw3
3 BR92 Dw3Dw3 15 7050A/NK1 dw3dw3
4 3535 dw3dw3 16 7050A/3535 dw3dw3
5 0-01 dw3dw3 17 01-26A dw3dw3
6 NK1 dw3dw3 18 01-26A/0-01 dw3dw3
7 LTR168 Dw3Dw3 19 01-26A/NK1 dw3dw3
8 7050A dw3dw3 20 01-26A/3550 Dw3dw3
9 7050A/LTR168 Dw3dw3 21 01-26A/BR92 Dw3Dw3
10 01-26A/LTR168 Dw3dw3 22 01-26A/3535 dw3dw3
11 7050A/0-01 dw3dw3 23 01-26A/LNR-4 Dw3dw3
12 7050A/3550 Dw3dw3

Table 9

Dwarf genotypes and their average plant height"

品系或组合Line or combination 基因型Genotype 平均株高Average plant height (cm)
LNR-4 Dw1Dw1Dw2Dw2Dw3Dw3 145.3
3550 dw1dw1Dw2Dw2Dw3Dw3 164.8
BR92 Dw1Dw1Dw2Dw2Dw3Dw3 162.4
3535 Dw1Dw1Dw2Dw2dw3dw3 145.5
0-01 Dw1Dw1dw2dw2dw3dw3 139.2
NK1 Dw1Dw1Dw2Dw2dw3dw3 139.1
LTR168 Dw1Dw1Dw2Dw2Dw3Dw3 247.3
7050A Dw1Dw1Dw2Dw2dw3dw3 136.7
7050A/LTR168 Dw1Dw1Dw2Dw2Dw3dw3 355.3
01-26A/LTR168 Dw1dw1Dw2Dw2Dw3dw3 341.7
7050A/0-01 Dw1Dw1Dw2dw2dw3dw3 178.4
7050A/3550 Dw1dw1Dw2Dw2Dw3dw3 185.7
7050A/LNR-4 Dw1Dw1Dw2Dw2Dw3dw3 173.1
7050A/BR92 Dw1Dw1Dw2Dw2Dw3dw3 179.2
7050A/NK1 Dw1Dw1Dw2Dw2dw3dw3 177.6
7050A/3535 Dw1Dw1Dw2Dw2dw3dw3 159.6
01-26A dw1dw1Dw2Dw2dw3dw3 103.5
01-26A/0-01 Dw1dw1Dw2dw2dw3dw3 141.9
01-26A/NK1 Dw1dw1Dw2Dw2dw3dw3 159.7
01-26A/3550 dw1dw1Dw2Dw2Dw3dw3 152.5
01-26A/BR92 Dw1dw1Dw2Dw2Dw3Dw3 146.6
01-26A/3535 Dw1dw1Dw2Dw2dw3dw3 144.8
01-26A/LNR-4 Dw1dw1Dw2Dw2Dw3dw3 141.3
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