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Acta Agronomica Sinica ›› 2024, Vol. 50 ›› Issue (2): 310-324.doi: 10.3724/SP.J.1006.2023.34066

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS • Previous Articles     Next Articles

Association analysis of three breeding traits with SSR markers and exploration of elite alleles in sugarcane

TIAN Chun-Yan1,2(), BIAN Xin1,2, LANG Rong-Bin1,2, YU Hua-Xian1,2, TAO Lian-An1,2, AN Ru-Dong1,2, DONG Li-Hua1,2, ZHANG Yu1,2, JING Yan-Fen1,2,*()   

  1. 1Academy of Agricultural Sciences / Yunnan Key Laboratory of Sugarcane Genetic Improvement, Kaiyuan 661699, Yunnan, China
    2Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of Agriculture and Rural Affairs, China (Yunnan), Kaiyuan 661699, Yunnan, China
  • Received:2023-03-30 Accepted:2023-06-29 Online:2024-02-12 Published:2023-07-19
  • Contact: *E-mail: rljyf@126.com
  • Supported by:
    Yunnan Seed Laboratory Project(202205AR070001-09);National Natural Science Foundation of China(31860406);Yunnan Fundamental Research Projects(2019FB053)

Abstract:

Cane yield and sucrose content are the two main breeding objectives in sugarcane, and plant height, stem diameter, and brix are major constituent factors of cane yield and sucrose. Identification of associated molecular markers and exploring elite alleles with yield and sucrose traits could provide a theoretical basis and reference for marker assisted breeding of sugarcane. In this study, 62 sugarcane germplasm entries were used as the experimental materials. Three breeding traits, including plant height, stem diameter, and brix were investigated under four planting environments at maturity stage. ANOVA, genetic variation analysis and association analysis between the identified from related markers for further allele effect analysis. The result of ANOVA revealed that genotype, planting environment and their interaction had significant effect on plant height, stem diameter and brix in sugarcane. The broad-sense heritability of the three traits ranged from 0.68 to 0.76, indicating that they had stable genetic characteristics and their phenotype was mainly determined by its genotype. The frequency of these three traits showed typical quantitative traits with normal distribution characteristics. The coefficients of variation of the three traits ranged from 6.73% to 19.89%, suggesting a rich variation in phenotypes. A total of 204 alleles were found with a mean of 5.5135 per marker. The average gene diversity coefficient was 0.6779, and the average PIC value was 0.6252. The 89.19% of total markers were highly polymorphic with PIC>0.5. Population structure analysis revealed that the tested group could be divided into three subgroups. A total of 20 markers were detected associated with plant height, stem diameter, and brix traits based on MLM method, which explained 5.04%-27.98% of phenotypic variation. Seven of them detected in two or more planting environments were considered as elite markers, 8 alleles with positive effect were identified and considered as the elite alleles, and 10 typical materials were found carrying these elite alleles by allele effect analysis. These results provide a great significance for candidate gene mining of yield and sucrose content traits and parents selection of hybrid combinations in sugarcane breeding.

Key words: sugarcane, breeding traits, SSR, association analysis, elite allele, allele effect

Table S1

Tested materials used in this study"

编号
Number
材料名称
Materials name
类型
Type
1 云瑞15-468 Yunrui 15-468 创新种质Innovation germplasm
2 云瑞17-142 Yunrui 17-142 创新种质Innovation germplasm
3 云瑞15-358 Yunrui 15-358 创新种质Innovation germplasm
4 云瑞17-135 Yunrui 17-135 创新种质Innovation germplasm
5 云瑞05-292 Yunrui 05-292 品系/亲本Line/parent
6 云瑞05-770 Yunrui 05-770 品系/亲本Line/parent
7 云瑞14-179 Yunrui 14-179 创新种质Innovation germplasm
8 云瑞14-177 Yunrui 14-177 创新种质Innovation germplasm
9 云瑞14-199 Yunrui 14-199 创新种质Innovation germplasm
10 云瑞17-146 Yunrui 17-146 创新种质Innovation germplasm
11 云瑞15-78 Yunrui 15-78 创新种质Innovation germplasm
12 云瑞03-315 Yunrui 03-315 品系/亲本Line/parent
13 云瑞17-148 Yunrui 17-148 创新种质Innovation germplasm
14 云瑞08-254 Yunrui 08-254 品系/亲本Line/parent
15 云瑞15-55 Yunrui 15-55 创新种质Innovation germplasm
16 云瑞17-133 Yunrui 17-133 创新种质Innovation germplasm
17 云瑞17-124 Yunrui 17-124 创新种质Innovation germplasm
18 云瑞15-566 Yunrui 15-566 创新种质Innovation germplasm
19 云瑞11-256 Yunrui 11-256 创新种质Innovation germplasm
20 云瑞17-114 Yunrui 17-114 创新种质Innovation germplasm
21 云瑞17-149 Yunrui 17-149 创新种质Innovation germplasm
22 云瑞15-503 Yunrui 15-503 创新种质Innovation germplasm
23 云瑞17-111 Yunrui 17-111 创新种质Innovation germplasm
24 云瑞15-56 Yunrui 15-56 创新种质Innovation germplasm
25 云瑞16-3 Yunrui 16-3 创新种质Innovation germplasm
26 云瑞16-4 Yunrui 16-4 创新种质Innovation germplasm
27 云瑞16 Yunrui 16 创新种质Innovation germplasm
28 云瑞37 Yunrui 37 创新种质Innovation germplasm
29 云瑞46 Yunrui 46 创新种质Innovation germplasm
30 云瑞52 Yunrui 52 创新种质Innovation germplasm
31 云瑞90 Yunrui 90 创新种质Innovation germplasm
32 云瑞151 Yunrui 151 创新种质Innovation germplasm
33 云瑞154 Yunrui 154 创新种质Innovation germplasm
34 云瑞164 Yunrui 164 创新种质Innovation germplasm
35 云瑞215 Yunrui 215 创新种质Innovation germplasm
36 云瑞258 Yunrui 258 创新种质Innovation germplasm
37 云蔗05-51 Yunzhe 05-51 品种/亲本Variety/parent
38 云蔗08-1609 Yunzhe 08-1609 品种/亲本Variety/parent
39 新台糖22号 ROC22 品种/亲本Variety/parent
40 新台糖16号 ROC16 品种/亲本Variety/parent
41 新台糖10号 ROC10 品种/亲本Variety/parent
42 粤糖60号 Yuetang 60 品种/亲本Variety/parent
43 粤糖93-159 Yuetang 93-159 品种/亲本Variety/parent
44 粤糖86-368 Yuetang 86-368 品种/亲本Variety/parent
45 粤糖00-236 Yuetang 00-236 品种/亲本Variety/parent
46 桂糖11号 Guitang 11 品种/亲本Variety/parent
47 赣蔗8号 Ganzhe 8 品种/亲本Variety/parent
48 柳城05-136 Liucheng 05-136 品种/亲本Variety/parent
49 福农15号 Funong 15 品种/亲本Variety/parent
50 福农39号 Funong 39 品种/亲本Variety/parent
51 崖城58-47 Yacheng 58-47 亲本Parent
52 崖城97-47 Yacheng 97-47 亲本Parent
53 园林6号 Yuanlin 6 品种/亲本Variety/parent
54 POJ3016 品种/亲本Variety/parent
55 POJ213 品种/亲本Variety/parent
56 Co475 品种/亲本Variety/parent
57 Q171 品种/亲本Variety/parent
58 Q208 品种/亲本Variety/parent
59 CP94-1196 品种/亲本Variety/parent
60 PK88-188 品种/亲本Variety/parent
61 VMC97-41 品种/亲本Variety/parent
62 SP81-3250 品种/亲本Variety/parent

Table 1

MANOVA and broad-sense heritability of three breeding traits"

性状
Trait
变异来源
Source of variation
自由度
DF
总方差
SS
均方
MS
P
P-value
广义遗传力
Broad-sense heritability
株高 PH 基因型 Genotype 61 525,860.4 8620.7 <0.001 0.68
环境 Environment 3 595,918.2 198,639.4 <0.001
基因型×环境 Genotype×Environment 183 321,306.8 1755.8 <0.001
茎径SD 基因型 Genotype 61 38.695,76 0.634,36 <0.001 0.76
环境 Environment 3 132.009,44 44.003,15 <0.001
基因型×环境 Genotype×Environment 183 13.438,21 0.073,43 <0.001
锤度Brix 基因型 Genotype 61 1383.048 22.673 <0.001 0.72
环境 Environment 3 386.028 128.676 <0.001
基因型×环境 Genotype×Environment 183 1007.080 5.503 <0.001

Table 2

Variation of three agronomic traits in tested sugarcane accessions under four planting environments"

性状
Trait
种植环境
Planting environment
平均值±标准差
Mean±SD
最大值
Maximum
最小值
Minimum
变异系数
CV (%)
偏度
Skewness
峰度
Kurtosis
株高
PH (cm)
FP_ND 263.48±28.25 324.73 197.60 10.72 -0.29 -0.41
FP_NM 264.40±28.63 330.44 205.00 10.83 0.10 -0.23
BP_RW 217.03±43.18 337.83 140.42 19.89 0.43 0.24
BP_DS 200.74±34.38 289.58 131.67 17.13 0.02 -0.51
茎径
SD (cm)
FP_ND 2.50±0.34 3.44 1.67 13.64 0.06 0.05
FP_NM 2.39±0.28 3.28 1.00 11.91 -0.38 1.76
BP_RW 1.58±0.23 2.00 0.89 14.89 -0.15 -0.65
BP_DS 1.62±0.21 2.06 1.11 12.98 -0.23 -0.32
锤度
Brix (%)
FP_ND 18.38±2.11 20.97 12.27 12.11 -1.49 1.89
FP_NM 18.95±1.27 22.21 15.56 6.73 0.05 0.13
BP_RW 17.97±1.92 22.42 13.93 10.71 0.19 -0.20
BP_DS 18.68±1.88 23.02 15.56 10.04 0.41 -0.55

Fig. 1

Frequency distribution of three breeding traits in 62 sugarcane germplasm entries"

Table 3

Polymorphism index of the 37 SSR marker pairs"

标记名称
Marker name
等位基因数
Alleles number
主要等位基因频率
Major allele frequency
多态性信息含量
PIC
基因多样性
Gene diversity
SMC334BS 9 0.3952 0.6605 0.7072
SMC336BS 9 0.2984 0.8129 0.8318
mSSCIR74 4 0.5000 0.5144 0.5951
mSSCIR3 7 0.2500 0.7961 0.8210
mSSCIR43 7 0.3790 0.6860 0.7288
SMC703BS 5 0.4435 0.6457 0.6951
SMC31CUQ 8 0.3387 0.7499 0.7802
SMC22DUQ 4 0.4435 0.5888 0.6554
SCB174 7 0.3145 0.7217 0.7607
SCB190 4 0.4194 0.6440 0.6976
DBF/Arb1 5 0.4194 0.6198 0.6778
DBF/Arb2 3 0.4758 0.4758 0.5704
DBF/Arb3 4 0.4919 0.4099 0.5235
Aqua/Arb1 2 0.5081 0.3749 0.4999
Aqua/Arb3 3 0.4355 0.5586 0.6353
SMC851MS 5 0.4113 0.6032 0.6645
SMC119CG 5 0.3387 0.6738 0.7248
SMC18SA 5 0.4113 0.6727 0.7172
SMC486CG 5 0.4435 0.6100 0.6700
SMC24DUQ 5 0.3145 0.7004 0.7453
SMC278CS 9 0.2661 0.8165 0.8362
SMC1604SA 5 0.5242 0.4876 0.5725
SMC7CUQ 4 0.5000 0.5599 0.6281
SMC1751CL 4 0.4194 0.5776 0.6457
SMC569CS 5 0.5242 0.5942 0.6424
SMC597CS 9 0.2500 0.7860 0.8124
mSSCIR66 5 0.3629 0.7107 0.7500
SMC36BUQ 5 0.3306 0.6590 0.7133
mSSCIR36 5 0.4677 0.6118 0.6677
SMC1825 4 0.5565 0.5665 0.6158
mSSCIR56 6 0.2984 0.7667 0.7964
SMC219 6 0.3710 0.6650 0.7158
SMC1752 5 0.4677 0.5913 0.6539
mSSCIR52 9 0.2500 0.8319 0.8490
IISR17b 4 0.3871 0.6698 0.7196
IISR111 3 0.4919 0.5528 0.6250
IISR306 10 0.2742 0.7918 0.8149
平均值 Mean 5.5135 0.4151 0.6252 0.6779

Fig. 2

Population structure of 62 sugarcane germplasm entries A: chart of ΔK changing with K-value; B: frequency distribution of Q-value in three populations; C: population structure of the 62 sugarcane accessions based on SSR data set."

Table 4

General overview of SSR markers associated with three breeding traits"

性状
Traits
关联位点个数
No. of associated loci
同时在2个及以上环境下检测到的位点数
No. of associated loci under two or more environments
表型变异解释率范围
Range of R2 (%)
株高Plant height 6 1 10.57-14.95
茎径Stem diameter 7 4 5.04-26.29
锤度Brix 7 2 16.90-27.98

Table 5

SSR markers associated with agronomic traits under two or more planting environments"

性状
Traits
标记
SSR markers
大田种植-南多
FP_ND
大田种植-弄门
FP_NM
桶栽-正常浇水
BP_RW
桶栽-干旱胁迫
BP_DS
株高 PH SCB190 - - 14.95** 10.57*
茎径 SD DBF/Arb1 15.18* 20.69* 16.37* -
SMC486CG 14.72** 26.29** - 13.47**
SMC1752 - 21.56** 12.40* -
IISR306 - 25.36** - 5.04*
锤度 Brix SMC851MS 16.90* 24.15* - -
SMC119CG 27.98** - - 26.50*

Table 6

Alleles of seven related elite markers and their phenotypic effect value"

性状
Trait
位点_等位变异
Locus_allele
表型效应Phenotypic effect of allele
FP_ND FP_NM BP_RW BP_DS
株高
PH (cm)
SCB190_247 bp - - -4.16 -2.81
SCB190_250 bp - - -7.64 -7.50
SCB190_253 bp - - -4.03 -1.59
SCB190_259 bp - - -0.16 -3.66
SCB190_263 bp - - +0.10 +2.02
茎径
SD (cm)
DBF/Arb1_217 bp +0.01 +0.02 +0.07 -
DBF/Arb1_221 bp +0.01 -0.01 +0.03 -
DBF/Arb1_222 bp -0.01 -0.02 +0.02 -
SMC486CG_222 bp +0.06 +0.06 - +0.06
SMC486CG_232 bp +0.02 +0.02 - +0.01
SMC486CG_234 bp -0.01 -0.01 - -0.01
SMC486CG_236 bp -0.07 -0.07 - -0.06
SMC486CG_237 bp -0.03 -0.06 - -0.04
SMC486CG_238 bp -0.18 -0.11 - -0.09
SMC486CG_242 bp -0.09 +0.03 - -0.02
SMC1752_135 bp - +0.02 -0.02 -
SMC1752_136 bp - -0.01 +0.01 -
SMC1752_138 bp - +0.01 -0.01 -
SMC1752_139 bp - -0.02 -0.03 -
IISR306_133 bp - -0.16 - -0.91
IISR306_134 bp - -0.13 - -0.92
IISR306_138 bp - -0.01 - -0.75
IISR306_139 bp - +0.15 - -0.75
IISR306_141 bp - +0.04 - -0.75
IISR306_142 bp - +0.04 - -0.75
IISR306_143 bp - +1.04 - -0.73
IISR306_144 bp - +2.04 - -0.73
锤度
Brix (%)
SMC851MS_129 bp -0.11 -0.08 - -
SMC851MS_131 bp -0.07 -0.09 - -
SMC851MS_132 bp -0.16 +0.11 - -
SMC851MS_133 bp +0.04 +0.01 - -
SMC851MS_134 bp +0.22 +0.04 - -
SMC851MS_137 bp -0.16 -0.26 - -
SMC119CG_110 bp +0.44 - - -0.09
SMC119CG_116 bp +0.40 - - +0.20
SMC119CG_122 bp +0.01 - - +0.51
SMC119CG_125 bp -5.27 - - -0.68
SMC119CG_132 bp -0.31 - - +0.12

Table 7

Excellent alleles and typical carrier materials in corresponding environments"

性状
Trait
位点_等位变异
Locus_allele
表型效应Phenotypic effect of allele 典型载体材料及其表型值
Typical carrier material and its phenotypic value
FP_ND FP_NM BP_RW BP_DS
株高
PH (cm)
SCB190_263bp - - +0.10 +2.02 云瑞164 (337.83 cm) Yunrui 164 (337.83 cm)
茎径
SD (cm)
DBF/Arb1_217bp +0.01 +0.02 +0.07 - 云瑞15-566 (3.44 cm), 粤糖86-368 (2.93 cm), 粤糖60号(2.92 cm)
Yunrui 15-566 (3.44 cm), Yuetang 86-368
(2.93 cm), Yuetang 60 (2.92 cm)
SMC486CG_222bp +0.06 +0.06 - - 云蔗08-1609 (3.09 cm), 粤糖86-368 (2.93 cm), 粤糖93-159 (2.52 cm)
Yunzhe 08-1609 (3.09 cm), Yuetang 86-368
(2.93 cm), Yuetang 93-159 (2.52 cm)
SMC486CG_232bp +0.02 +0.02 - +0.01 云蔗08-1609 (3.09 cm), 桂糖11号 (3.28 cm),
粤糖93-159 (2.52 cm)
Yunzhe 08-1609 (3.09 cm), Guitang 11 (3.28 cm), Yuetang 93-159 (2.52 cm)
锤度
Brix (%)
SMC851MS_133bp +0.04 +0.01 - - 云蔗08-1609 (22.97%), 福农15号 (22.21%)
Yunzhe 08-1609 (22.97%), Funong 15 (22.21%)
SMC851MS_134bp +0.22 +0.04 - - 云蔗08-1609 (22.97%), 福农15号 (22.21%)
Yunzhe 08-1609 (22.97%), Funong 15 (22.21%)
SMC119CG_116bp +0.40 - - +0.20 云蔗08-1609 (22.97%), 粤糖00-236 (23.02%)
Yunzhe 08-1609 (22.97%), Yuetang 00-236 (23.02%)
SMC119CG_122bp +0.01 - - +0.51 Co475 (20.39%), 粤糖00-236 (23.02%)
Co475 (20.39%), Yuetang 00-236 (23.02%)
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