Scientia Agricultura Sinica ›› 2022, Vol. 55 ›› Issue (22): 4342-4355.doi: 10.3864/j.issn.0578-1752.2022.22.002

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

Construction of A Comprehensive Evaluation System and Screening of Cold Tolerance Indicators for Cold Tolerance of Cotton at Seedling Emergence Stage

SHEN Qian1,2(),ZHANG SiPing1,LIU RuiHua1,LIU ShaoDong1,CHEN Jing1,GE ChangWei1,MA HuiJuan1,ZHAO XinHua1,YANG GuoZheng2,SONG MeiZhen1(),PANG ChaoYou1()   

  1. 1Institute of Cotton, Chinese Academy of Agricultural Sciences/State Key Laboratory of Cotton Biology, Anyang 455000, Henan
    2College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070
  • Received:2022-07-13 Accepted:2022-08-15 Online:2022-11-16 Published:2022-12-14
  • Contact: MeiZhen SONG,ChaoYou PANG E-mail:shenqian429@126.com;songmzccri@163.com;chypang@163.com

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Abstract:

【Objective】 In this study, the purpose was to comprehensively evaluate the cold tolerance of cotton varieties (lines) at seedling emergence stage, establish a reliable evaluation model, screen and identify indicators, and provide a simple and effective evaluation method for the selection and identification of cold-tolerant varieties in cotton.【Method】200 upland cotton varieties (lines) were used to test hypocotyl length, root length and 100-grain weight, etc. under three treatments of constant chilling (CC), diurnal variation of chilling (DVC) and normal conditions. A combination of integrated cold tolerance coefficient difference analysis, frequency analysis, drop analysis, principal component analysis, cluster analysis, and multiple regression analysis were used to classify their cold tolerance types, establish cold tolerance prediction models, and screen evaluation parameters. 【Result】The variation of each parameters at normal conditions were minor fluctuations ranging from 3.12% to 18.89%. The seedling emergence rate was above 85.00%, which had high viability and could be used for subsequent cold tolerance analysis. The variability of each parameter within the accessions increased under chilling stress, ranging from 7.14%-108.33%, and the most variable parameter were root length under CC condition and germination index under DVC condition. Principal component analysis converted the 14 parameters under chilling stress and 100-grain weight measured into six mutually independent composite indicators, representing 74.98% of the total data information. The comprehensive cold tolerance evaluation value (D) was calculated by the affiliation function method and then clustering analysis was performed. 200 cotton varieties (lines) were divided into five categories according to their cold tolerance, with 2 of the group Ⅰ being strongly cold tolerant, 42 of the group Ⅱ being cold tolerant, 69 of the group Ⅲ being medium cold tolerant, 83 of the group Ⅳ being more sensitive, and 4 of the group Ⅴ being sensitive, of which Xinluzhong 16 was the most cold-tolerant material. A multiple regression analysis was applied to establish a prediction model for cold tolerance of cotton at seedling emergence as Y=-4.10+0.58X4+0.40X14+0.32X1+0.22X5 (R2=0.92), and four parameters for cold resistance evaluation were confirmed, namely total length of seedling, emergence rate, and dry weight under CC stress, germination rate under DVC stress. The cold-tolerant varieties (lines) had higher seedling emergence rates of early sowing experiment in the field, which were basically consistent with the results of the indoor results. 【Conclusion】 It is feasible to use CC and DVC stress combined with multivariate statistical analysis to evaluate the cold tolerance of cotton at seeding stage, and total length of seedling, emergence rate, and dry weight under CC stress, germination rate under DVC stress can be used as evaluation parameters.

Key words: Gossypium hirsutum L., seedling emergence stage, cold tolerance, comprehensive evaluation, parameters screening

Fig. 1

Matrix scatter analysis of basic traits of 200 cotton varieties (lines) HGW: 100 grain weight; ER: Emergence rate; HL: Hypocotyl length; RL: Root length; TL: Total length of seedling; GR: Germination rate; GI: Germination index; DW: Dry weight; SS: Soluble sugar content; SAA: Soluble amino acid content. * and ** mean significantly different at the 0.05 and 0.01 probability levels, respectively. The same as below"

Table 1

Statistical analysis on basic traits of 200 cotton varieties (lines)"

指标
Parameter		 均值
Mean		 中位数
Median		 变异系数
Coefficient variation		 标准差
Standard deviation		 最小值
Minimum		 最大值
Maximum		 偏度
Skewness		 峰度
Kurtosis
百粒重HGW (g) 9.71 9.50 11.12 1.08 7.13 12.98 0.41 0.07
出苗率ER (%) 94.34 95.00 6.53 6.16 85.00 100.00 -0.84 -0.36
下胚轴长度HL (cm) 10.11 10.10 6.82 0.69 8.20 12.68 0.14 0.52
根长RL (cm) 12.14 12.20 10.63 1.29 8.27 15.81 -0.32 0.22
总长TL (cm) 22.24 22.30 7.46 1.66 16.60 26.85 -0.2 0.24
萌发率GR (%) 99.05 100.00 3.12 3.09 60.00 100.00 -6.43 61.68
萌发指数GI 14.72 14.50 18.89 2.78 7.00 20.00 0.11 -0.95
干物重DW (mg/plant) 70.32 69.00 14.60 10.27 46.00 103.90 0.45 -0.07
可溶性糖含量SS (mg·g-1) 49.92 47.45 12.70 6.34 32.14 66.50 0.15 0.035
游离氨基酸含量SAA (mg·g-1) 28.12 27.65 17.99 5.06 14.73 40.30 -0.31 -0.07

Table 2

Difference analysis of germination rate of 20 cotton varieties after chilling treatment for different time"

低温处理
Chilling stress		 时间
Time (d)		 最小值
Minimum (%)		 最大值
Maximum (%)		 平均值
Mean		 变异系数
Coefficient variation (%)
恒定低温
Constant chilling		 1 59.26 99.97 85.89 17.02
3 36.19 98.25 69.81 37.29
5 0.00 68.03 40.77 88.04
7 0.00 15.56 2.84 202.56
9 0.00 24.44 2.19 265.24
昼夜变温
Diurnal variation of chilling		 10 13.73 100.00 66.22 47.52
12 20.95 96.30 60.51 50.29
13 0.00 90.04 43.38 107.41
14 0.00 84.44 23.02 126.45
16 0.00 59.26 23.23 123.32

Table 3

Difference analysis of parameters of cotton materials at seedling emergence stage under chilling stress"

CC：恒定低温；DVC：昼夜变温。下同

CC: Constant chilling; DVC: Diurnal variation of chilling. The same as below

指标
Parameter		 均值
Mean		 中位数
Median		 变异系数
Coefficient variation
(%)		 标准差
Standard deviation		 最小值
Minimum		 最大值
Maximum		 偏度
Skewness		 峰度
Kurtosis
ER-CC 0.66 0.67 21.21 0.14 0.00 0.99 -0.32 -0.25
HL-CC 0.70 0.70 7.14 0.05 0.52 0.83 -0.16 0.43
RL-CC 0.46 0.44 21.74 0.10 0.31 0.94 1.76 4.71
TL-CC 0.58 0.57 10.34 0.06 0.42 0.77 0.57 0.56
WD-CC 0.74 0.74 10.81 0.08 0.56 0.93 0.21 -0.09
SAA-CC 0.85 0.89 23.53 0.20 0.30 1.36 -0.41 -0.35
SS-CC 1.24 1.23 11.29 0.14 0.79 1.95 0.88 3.84
ER-DVC 0.63 0.64 25.40 0.16 0.16 0.96 -0.34 -0.06
HL-DVC 0.78 0.79 7.69 0.06 0.65 0.89 -0.15 0.29
RL-DVC 0.66 0.65 19.70 0.13 0.41 1.12 0.57 1.80
TL-DVC 0.72 0.71 11.11 0.08 0.56 1.00 0.51 1.51
WD-DVC 0.73 0.73 12.33 0.09 0.51 0.92 0.05 -0.05
GI-DVC 0.35 0.32 62.86 0.22 0.00 0.63 2.11 4.38
GR-DVC 0.12 0.08 108.33 0.13 0.02 0.81 0.50 -0.41

Table 4

The cold resistance coefficients of the cotton materials and their distribution in different intervals"

指标
Parameter		 0<CTC≤0.2 0.2<CTC≤0.4 0.4<CTC≤0.6 0.6<CTC≤0.8 0.8<CTC≤1.2 1.2<CTC≤1.8
次数
Times		 频次
Freq. (%)		 次数
Times		 频次
Freq. (%)		 次数
Times		 频次
Freq. (%)		 次数
Times		 频次
Freq. (%)		 次数
Times		 频次
Freq. (%)		 次数
Times		 频次
Freq. (%)
ER-CC 38 19.00 40 20.00 52 26.00 41 20.50 29 14.50 0 0.00
HL-CC 0 0.00 0 0.00 7 3.50 188 94.00 5 2.50 0 0.00
RL-CC 0 0.00 54 27.00 132 66.00 12 6.00 2 1.00 0 0.00
TL-CC 0 0.00 137 68.50 63 31.50 0 0.00 0 0.00 0 0.00
DW-CC 0 0.00 0 0.00 6 3.00 148 74.00 46 23.00 0 0.00
SS-CC 0 0.00 0 0.00 0 0.00 1 0.50 74 37.00 125 62.50
SAA-CC 0 0.00 3 1.50 20 10.00 46 23.00 125 62.50 6 3.00
ER-DVC 2 1.00 13 6.50 68 34.00 90 45.00 27 13.50 0 0.00
HL-DVC 0 0.00 0 0.00 0 0.00 135 67.50 65 32.50 0 0.00
RL-DVC 0 0.00 0 0.00 55 27.50 130 65.00 15 7.50 0 0.00
TL-DVC 0 0.00 0 0.00 6 3.00 177 88.50 17 8.50 0 0.00
DW-DVC 0 0.00 0 0.00 10 5.00 154 77.00 36 18.00 0 0.00
GI-DVC 170 85.00 17 8.50 11 5.50 2 1.00 0 0.00 0 0.00
GR-DVC 46 23.00 87 43.50 43 21.50 23 11.50 1 0.50 0 0.00
CTC：耐冷系数。下同 CTC: Cold tolerance coefficient. The same as below

Fig. 2

Effects of chilling damage on each parameter at seeding stage of cotton"

Table 5

Power vector (PV), characteristic value, contribution rate and cumulative contribution rate of six principal components"

性状
Trait		 特征向量Power vector
PV1 PV2 PV3 PV4 PV5 PV6
ER-CC 0.52 0.00 0.11 0.42 0.35 0.05
HL-CC 0.87 0.17 -0.02 -0.04 -0.24 0.09
RL-CC 0.81 0.20 0.01 -0.25 0.045 -0.09
TL-CC 0.94 0.23 -0.02 -0.14 -0.11 0.00
DW-CC 0.69 0.15 0.17 -0.19 -0.08 0.06
SS-CC 0.31 0.10 -0.09 0.56 0.13 -0.07
SAA-CC -0.30 0.07 0.39 -0.27 -0.33 -0.37
ER-DVC -0.10 0.46 -0.25 0.58 -0.24 0.13
HL-DVC -0.10 0.65 -0.25 0.08 -0.15 0.05
RL-DVC -0.23 0.77 -0.36 -0.22 0.07 -0.03
TL-DVC -0.23 0.85 -0.36 -0.21 0.03 -0.01
DW-DVC -0.11 0.28 0.09 -0.17 0.82 -0.13
GI-DVC -0.12 0.50 0.80 0.07 -0.02 0.12
GR-DVC -0.18 0.55 0.73 0.17 -0.02 0.05
HGW -0.15 -0.09 0.04 -0.25 0.06 0.89
特征值 Characteristic value 3.40 2.74 1.76 1.24 1.10 1.01
方差贡献率 Contribution rate (%) 22.65 18.25 11.74 8.26 7.35 6.72
累积贡献率
Cumulative contribution rate (%)		 22.65 40.90 52.64 60.90 68.25 74.98

Fig. 3

Cold tolerance evaluation of cotton varieties (lines) based on D value A: Cold tolerance phylogenetic map; B: Geographical distribution of varieties (lines) in five groups. FR: Foreign regions; NEM: Northern early maturity; NW: Northwest inland region; YR: Yellow River region; YZR: Yangtze River region"

Table 6

Prediction model of cold tolerance of cotton varieties (lines)"

X1：恒定低温下出苗率；X4：恒定低温下的棉苗总长；X5：恒定低温下的干物重；X6：恒定低温下的可溶性糖含量；X14：昼夜变温下的萌发率

X1: ER-CC; X4: TL-CC; X5: DW-CC; X6: SS-CC; X14: GR-DVC

多元回归方程
Multiple regression equation		 相关系数
r		 判定系数
R2		 F值
F value		 P值
P value
Y=-3.28+0.80X4 0.80 0.64 342.76 ≤0.001
Y=-3.94+0.82X4+0.42X14 0.90 0.81 409.48 ≤0.001
Y=-3.84+0.71X4+0.41X14+0.31X1 0.94 0.87 506.66 ≤0.001
Y=-4.10+0.58X4+0.40X14+0.32X1+0.22X5 0.96 0.92 537.36 ≤0.001
Y=-5.02+0.54X4+0.39X14+0.29X1+0.23X5+0.17X6 0.97 0.94 654.30 ≤0.001

Fig. 4

The effect of early sowing in the field on the germplasm emergence rate A: Emergence rate in Field; B: Correlation analysis between the comprehensive value of varieties (lines) obtained under indoor conditions and field emergence rate; D: The comprehensive value of each parameter under CC and DVC stress; D-CC: The comprehensive value of each parameter under CC stress; D-DVC: The comprehensive value of each parameter under DVC stress; ER-2021: The emergence rate of the tested material in 2021; ER-2022: The emergence rate of the tested material in 2022; C: Field temperature in 2021; D: Field temperature in 2022; Date (month/day)"

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