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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (9): 2454-2466.doi: 10.3724/SP.J.1006.2025.51016

• TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY • Previous Articles     Next Articles

Comparative analysis of metabolite changes during young panicle development in wheat AL type cytoplasmic male serile line and homologous maintainers

KONG De-Zhen(), SANG Wei, NIE Ying-Bin, LI Wei, XU Hong-Jun, LI Jiang-Bo, LIU Peng-Peng, TIAN Xiao-Ming*()   

  1. Institute of Crop Research, Xinjiang Academy of Agri-Reclamation Sciences / Key Lab of Xinjiang Production and Construction Corps for Cereal Quality Research and Genetic Improvement, Shihezi 832000, Xinjiang, China
  • Received:2025-02-18 Accepted:2025-06-01 Online:2025-09-12 Published:2025-06-20
  • Contact: *E-mail: txming729@vip.sina.com E-mail:kongdezhen1746@163.com;txming729@vip.sina.com
  • Supported by:
    Xinjiang Production and Construction Corps Financial Science and Technology Plan Project(2023CB007-10);Xinjiang Production and Construction Corps Financial Science and Technology Plan Project(2024DA022)

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Abstract:

To investigate the relationship between the AL-type cytoplasmic male sterile (CMS) wheat line and the metabolome of its maintainer line during spikelet development, and to provide a theoretical foundation for the use of exogenous hormones in regulating fertility restoration, the AL-22A sterile line and the AL-22B homotypic maintainer line were used as experimental materials. Self-seed setting rate was determined using international standard methods. Spike samples were collected at nine developmental stages: young panicle differentiation, double ridge, floret primordium differentiation, pistil and stamen differentiation, chamber formation, tetrad, mononuclear median, uninucleate, binucleate, and trinucleate stages. The levels of endogenous hormones, peroxidase (POD), reactive oxygen species (ROS), and ATP were measured using liquid chromatography-mass spectrometry (LC-MS), and metabolites were analyzed through non-targeted metabolomics. The self-seed setting rate of the sterile line was zero, indicating complete sterility. Compared to the maintainer line, the sterile line showed higher levels of abscisic acid (ABA) and lower levels of indole-3-acetic acid (IAA), zeatin riboside (ZR), and gibberellic acid (GA3). Methyl jasmonate (MeJA) content was lower in the sterile line from the double ridge to tetrad stages but higher after the mononuclear median stage. The IAA/ABA ratio was consistently lower in the sterile line, suggesting that hormone imbalance may contribute to sterility, particularly the elevated levels of ABA and MeJA. No significant differences in ATP and ROS levels were observed between the two lines across developmental stages. However, POD activity was significantly higher in the maintainer line from the double ridge to tetrad stages, while the sterile line showed distinct POD levels at the binucleate and trinucleate stages. Non-targeted metabolomic analysis revealed that the sterile line had more downregulated metabolites during the pistil and stamen differentiation and mononuclear median stages, while other stages showed more upregulated than downregulated metabolites, with significant differences. Clustering analysis of metabolite expression patterns across developmental stages grouped samples into early (I-III), middle (IV-VI), and late (VII-IX) phases. Stage-specific differences in metabolite expression were associated with fertility changes. Pathway enrichment analysis indicated that differential metabolites were primarily concentrated in the floret primordium differentiation, chamber formation, tetrad, uninucleate, and trinucleate stages, mainly involving carboxylic acid metabolism and secondary metabolism. The stages most critical to fertility development occurred in early to mid (II-IV) and mid to late (VI-VIII) phases. These results suggest that the interplay between endogenous hormone balance and abnormal metabolite accumulation may be key factors in CMS. This study provides both theoretical insights and practical references for understanding the physiological and metabolic mechanisms of spikelet development in CMS wheat and for developing strategies to promote fertility restoration in AL-type sterile lines.

Key words: wheat, AL-type, CMS, endogenous hormones, metabolome

Table 1

Specific sampling dates for each developmental stage"

系
Line		 发育时期Development stage (month/day) 自交结实率(国际法)
Self-seeds rate (international, %)
I II III IV V VI VII VIII IX
不育系 Sterile line (A) 04/09 04/19 04/24 04/28 05/03 05/06 05/09 05/11 05/13 0
保持系 Maintainer line (B) 04/09 04/19 04/24 04/28 05/03 05/06 05/09 05/11 05/13 100

Fig. S1

Quantitative standard curves of five hormone external standard methods"

Fig. 1

Changes in endogenous hormone levels in male sterile and maintainer lines at different developmental stages Abbreviations are the same as those given in Table 1. A: sterile line; B: maintenance line. ABA: abscisic acid; ZR: zeatin riboside; MeJA: methyl jasmonate; IAA: indole-3-acetic acid; GA3: gibberellin acid. Different lowercase letters mean significant difference at 5% levels."

Table 2

Ratio of endogenous hormone levels between male sterile and maintainer lines"

项目Item 材料Material I II III IV V VI VII VIII IX
IAA/ABA A 0.50 1.17 0.40 0.35 0.26 0.29 2.69 0.69 2.89
B 0.97 2.21 0.49 0.94 0.66 0.69 3.87 3.53 4.13
B-A 0.47 1.04 0.09 0.59 0.40 0.40 1.18 2.84 1.24
IAA/GA3 A 5.09 41.69 16.79 44.18 29.20 21.30 43.46 1.25 4.57
B 6.71 24.16 71.28 29.73 24.89 1.99 3.81 5.86 129.94
B-A 1.62 -17.53 54.49 -14.45 -4.31 -19.31 -39.65 4.61 125.37
ABA/GA3 A 10.12 35.65 42.27 125.45 111.59 73.46 16.17 1.81 1.58
B 6.94 10.92 145.97 31.51 37.96 2.87 0.98 1.66 31.43
B-A -3.18 -24.73 103.70 -93.94 -73.63 -70.59 -15.19 -0.15 29.85
MeJA/IAA A 17.19 3.03 3.83 1.00 1.46 71.66 40.08 18.87 0.98
B 21.98 21.39 8.39 5.33 9.95 4.05 3.82 2.00 1.76
B-A 4.79 18.36 4.56 4.33 8.49 -67.61 -36.26 -16.87 0.78
ZR/ IAA A 28.89 14.62 18.57 9.60 12.86 45.00 15.03 24.42 8.93
B 16.50 17.33 13.54 11.16 8.02 20.95 12.84 8.38 7.34
B-A -12.39 2.71 -5.03 1.56 -4.84 -24.05 -2.19 -16.04 -1.59
ZR/ GA3 A 147.13 609.41 311.74 423.92 375.51 958.56 653.39 30.51 40.84
B 110.67 418.64 965.37 331.77 199.58 41.58 48.91 49.09 953.25
B-A -36.46 -190.77 653.63 -92.15 -175.93 -916.98 -604.48 18.58 912.41
MeJA/ GA3 A 87.57 126.52 64.36 44.29 42.65 1526.45 1742.01 23.58 4.50
B 147.38 516.89 598.18 158.33 247.66 8.04 14.54 11.72 228.15
B-A 59.81 390.37 533.82 114.04 205.01 -1518.41 -1727.47 -11.86 223.65

Fig. 2

Changes of energy matter and enzyme activity of sterile line and maintainer line in different stages of wheat development Abbreviations are the same as those given in Table 1. A: sterile line; B: maintenance line. Different lowercase letters mean significant differences at the 5% levels level (P < 0.05)."

Fig. 3

Analysis of metabolites difference between sterile line and maintainer line in different stages of wheat development Abbreviations are the same as those given in Table 1."

Fig. 4

Different metabolites enriched by sterile line and maintainer line in the first ten stages of spikelet development"

Fig. 5

KEGG pathway analysis of different metabolites in sterile and maintainer lines at different periods Abbreviations are the same as those given in Table 1."

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