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Acta Agronomica Sinica ›› 2023, Vol. 49 ›› Issue (1): 62-72.doi: 10.3724/SP.J.1006.2023.24007

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Analysis of key genes involved in GA pathway responding to temperature and exogenous GA related to internode development in soybean

QI Yang-Yang(), DOU Ru-Na, ZHAO Cai-Tong, ZHANG Zhi, LI Wen-Bin, JIANG Zhen-Feng()   

  1. Northeast Agricultural University, Key Laboratory of Soybean Biology in Chinese Ministry of Education / Key Laboratory of Soybean Biology and Genetics Breeding of Ministry of Agriculture and Rural Affairs, Harbin 150030, Heilongjiang, China
  • Received:2022-01-04 Accepted:2022-03-25 Online:2023-01-12 Published:2022-04-20
  • Contact: JIANG Zhen-Feng E-mail:qyy2022@126.com;jzhf@neau.edu.cn
  • Supported by:
    National Natural Science Foundation of China(31571693);National Natural Science Foundation of China(32172072);China Agriculture Research System of MOF and MARA (Soybean, Key Task)(CARS-04-04B);Natural Science Foundation of Heilongjiang Province(LH2021C025)

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Abstract:

The objective of this study is to explore the effects of temperature and exogenous GA on the development of soybean internode and the key genes related to gibberellin (GA) signal transduction pathway. The soybean variety ‘Charleston’ was grown in chambers or in pots under outdoor condition, and subjected to the treatments with different temperature and concentration of GA solutions. Phenotype observation, section with hand, LC-MS analysis, and RNA-seq experiments were performed to investigate the internode changes and the genes involved in the GA signal transduction pathway. Different temperature and exogenous GA all induced the internode elongation and the longer the length increased, the slender the internode became. Exogenous GA had an evident elongating effect on the cell length of the internode despite of no effect on the cell width. The internodes grown in 30℃ growth condition were longer than those in 25℃ growth condition. GA2-oxidase, GA19, GA53, GA20, and bioactive GA3 were detected in the elongation zone of soybean internode, suggesting that GA2-oxidase might play an important role in the developmental process of soybean internode. Tissue-specific genes in DELLA, GAI, and PIF gene families could be identified and facilitate the gene selection to regulate the internode growth and plant height from GA signal transduction pathway. Soybean internode was regulated by temperature and exogenous GA. From GA53 to GA19 to GA20 and ultimately to GA3 was an important GA synthesis pathway. GA2-oxidase played an important role on the internode development of soybean. The GA-GID1-DELLA complex was expressed specifically in soybean internode. The candidate genes related to the internode development could be filtered from GID1 and DELLA gene family.

Key words: soybean, internode development, GA pathway, GA-GID1-DELLA

Fig. 1

High temperature accelerates the internode development of soybean plant Left: two plants grown at 30℃; Right: two plants grown at 25℃."

Table 1

Effect of vary temperature on the developmental internode of soybean plants"

温度处理
Temperature treatment		 处理前
Before treatment (cm)		 12 h后
After 12 hours (cm)		 差值
Range (cm)
30℃ 3.02±0.32 3.38±0.35 0.36±0.03
25℃ 2.74±0.13 2.99±0.14 0.25±0.01

Fig. 2

Exogenous GA accelerates the growth of soybean internode A: 100 mmol L-1 GA treatment; B: 250 mmol L-1 GA treatment; C: CK."

Fig. 3

Effects of different concentrations of exogenous GA on the growth of soybean internodes A: the internode length of GA treatment at 25℃; B: the internode length of GA treatment at 30℃. 0 h: the internode length before GA treatment; 24 h: the internode length 24 hours after GA treatment."

Fig. 4

Effects of exogenous GA on the cells of elongation zone (EZ) and mature zone (MZ) in the soybean developmental internode in vary temperatures A: EZ grown in 30℃; B: MZ grown at 30℃; C: EZ grown in 25℃; D: MZ grown at 25℃."

Table 2

Cell length and width of soybean developmental internode in vary temperatures with exogenous GA"

节间
Internode		 30℃ 25℃
长度 Length (pixel) 宽度 Width (pixel) 长度 Length (pixel) 宽度 Width (pixel)
伸长区 EZ 206.00±4.48 63.78±6.00 131.59±6.48 54.87±1.20
成熟区 MZ 708.33±21.20 65.78±1.11 665.79±27.51 79.18±2.51

Table 3

GA content in the developmental internode of soybean plants (ng g-1)"

节间
Internode		 GA3 GA15 GA19 GA20 GA53
伸长区 EZ 0.13±0.01 0.24±0.03 48.14±0.01 0.24±0.02 19.40±0.01
成熟区 MZ — — 47.91±0.02 — 19.38±0.02

Fig. 5

Relative expression levels of key genes involved in internode development of soybean plant grown in vary temperature and exogenous GA treatments HCK: control at 30℃; LCK: control at 25℃; HG: GA treatment at 30℃; LG: GA treatment at 25℃."

Fig. 6

Relative expression levels of key genes related to GA under high or low plant densities CD1: the first internode of the apex in low plant density; CD21: the elongation zone of the second internode in low density; CD22: the mature zone of the second internode in low density; CD3: the third internode in low density; CH1: the first internode of the apex in high density; CH21: the elongation zone of the second internode in high density; CH22: the mature zone of the second internode in high density; CH3: the third internode in low density."

Table 4

Tissue-specific expression analysis of key genes related to GA signal transduction in soybean"

基因
Gene name		 嫩叶
Young leaf		 花
Flower		 1 cm荚
One-cm pod		 10 d种子
Seed of 10 DAF		 42 d种子
Seed of 42 DAF		 根
Root		 根瘤
Nodule
Glyma.12g213700
Glyma.13g288000 2.88 3.91 6.75
Glyma.20g141200 1.60 5.00 4.03
Glyma.05g130600 8.09
Glyma.13g361700 3.44 3.61 1.58 0.35 6.34 3.70
Glyma.15g012100 3.83 1.27 4.58 2.67 6.34 6.61
Glyma.04g150500 2.05 4.54 4.43 1.15 1.48 1.73 0.86
Glyma.06g213100 2.37 3.25 1.30 1.03 1.70 2.36 1.97
Glyma.06g213100 2.37 3.25 1.30 1.03 1.70 2.36 1.97
Glyma.11g216500 -2.88 -0.36 0.01 -0.03 -2.63 -2.33 -4.65
Glyma.18g040000 -2.31 -0.33 1.40 0.77 -3.97 -2.12 -4.03
Glyma.08g095800 -0.83 -1.01 -0.08 -0.87 -1.33 -1.96 -2.62
Glyma.20g200500 3.75 2.83 2.63 1.28 0.17 3.09 1.76
Glyma.05g140400 -1.51 -1.34 -0.84 -2.29 -1.65 -1.80 -1.94
Glyma.10g190200 3.66 3.14 2.00 1.03 -1.08 4.77 2.44
Glyma.15g141400 5.00 6.61
Glyma.03g148300 1.20 -1.75 0.65 -0.67 1.13 -1.49 -1.42
Glyma.10g022900 2.62 1.57 6.17 4.67 1.39 0.88 0.46
Glyma.02g151100 3.51 2.25 3.51 2.09 2.88 1.55 1.22
Glyma.20g230600 2.70 3.42 2.05 3.09 2.25 2.02 1.53
Glyma.10g158000 1.44 1.96 1.43 0.50 2.25 0.39 -0.10
Glyma.03g170300
Glyma.03g225000
Glyma.10g142600 3.66 2.54 3.85 -0.35 3.17 3.50 3.16
Glyma.20g091200 6.71 4.97 0.06
Glyma.02g282100 7.83 3.68 8.09
Glyma.U031209
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