Scientia Agricultura Sinica ›› 2026, Vol. 59 ›› Issue (5): 951-966.doi: 10.3864/j.issn.0578-1752.2026.05.003

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

Morphogenesis of the Low-Growth Point and Its Multi-Hormonal Regulatory Mechanism During Overwintering in Winter Rapeseed (Brassica napus L.)

LIU HaiQing1,2(), JIN JiaoJiao1,2, SUN WanCang3, CHAI Peng4, QI WeiLiang1,2, YANG Gang3, LI Chan1, LUO XueMei1, SU YunYun1,2, QIN XueXue1   

  1. 1 School of Agriculture and Bioengineering, Longdong Univerisity, Qingyang 745000, Gansu
    2 Gansu Key Laboratory of Protection and Utilization for Biological Resources and Ecological Restoration, Qingyang 745000, Gansu
    3 College of Agronomy, Gansu Agricultural University, Lanzhou 730070
    4 Xifeng Agricultural Technology Extension Center, Qingyang 745000, Gansu
  • Received:2025-08-19 Accepted:2025-11-12 Online:2026-03-01 Published:2026-03-06

Abstract:

【Objective】 This study aims to clarify the mechanism of plant hormones on the morphology of growth point and their response to cold resistance of Brassica napus, to provide a theoretical basis for breeding winter Brassica napus varieties, expanding its planting area, and increasing yield per unit.【Method】 Using 9 rapeseed varieties with distinct winter hardiness as experimental materials, a two year (2023-2025) consecutive field identification of overwintering rate was conducted in Qingyang, Gansu Province. Mid-November samplings were used to quantify shoot to root ratio, growth point height and additional phenotypic indicators of overwintering potential. Paraffin sectioning and trypan-blue/nitroblue tetrazolium staining were used to visualize growth cone morphology, cell death, and superoxide accumulation to evaluate the cold tolerance of different cultivars. Growth point of three representative genotypes that span the full range of cold hardiness (NTS309>Tianyou 14>Qinzao 1) were simultaneously analysed by targeted LC-MS/MS targeted phytohormone profiling and RNA-seq. PCA and O2PLS integrative analyses of the metabolome and transcriptome, the physiological and molecular mechanisms underlying the differences in growth point height among these varieties were systematically analyzed.【Result】 The results showed the following: Significant differences were observed among cultivars in overwintering rate, root collar diameter, dry weight, root/shoot ratio, and height of the growing point. Strong cold-tolerant varieties, less reactive oxygen species (ROS) and fewer dead cells observed following cold stress, accompanied by a lower proportion of cells damaged by low temperature, the opposite was true for weak cold-tolerant varieties. Statistical analysis revealed that the overwintering rate was extremely significantly and negatively correlated with growth point height (correlation coefficient r=-0.972, P<0.001). Rapeseed varieties with strong cold-tolerance (e.g., NTS309) which exhibited an overwintering rate exceeding 70%, had a relatively lower growth point height (approximately 3 cm), additionally, the height of the growth cone (observed in paraffin sections) was only 161.5 μm. In contrast, varieties with weak cold-tolerance (e.g., Qinzao 1) had growth points that protruded above the soil surface (approximately 5 cm), and their growth cone measured approximately 252.9 μm in height. Integrated analysis of hormone metabolomics and transcriptomics data revealed that the height of rapeseed growth point is regulated by a multi-dimensional regulatory network involving the phytohormones IAA, CKs, GA, JA and ABA. In strong cold-tolerant varieties, the levels of free IAA, conjugated IAA, JA-Ile, GA19, and ABA-GE were significantly up-regulated, whereas those of iP9G, mT9G, and BAP9G were significantly down-regulated. Differentially expressed genes (DEGs) between strong and weak cold-tolerant varieties were significantly enriched in pathways including “tryptophan metabolism”, “zeatin biosynthesis”, “plant hormone signal transduction”, and “α-linolenic acid metabolism”. The orthogonal partial least squares (O2PLS) model indicated that metabolites including IAA-Glu, tZR, and JA-Ile exhibited strong covariance with transcriptional modules, these interactions collectively form a hormone-transcription factor-metabolite feedback loop, which exerts three key regulatory effects: inhibiting cell elongation, promoting meristem dormancy and reducing the height of growth cone and growth points. Ultimately, this regulatory loop enhances the defense capacity of rapeseed against low-temperature freezing damage.【Conclusion】 This study is the first to confirm that the “dwarf growth point” of rapeseed is a key morphological marker for overwintering survival, and its formation is finely regulated by a multi-hormone network.

Key words: winter rapeseed (Brassica napus L.), plant hormone, growth point, growth cone, cold resistance

Table 1

Cultivar characteristics and origin"

序号No 品种(系)Varieties (Lines) 冬性Winterness 选育单位Source
1 GAU30 强冬性Strong winterness 甘肃农业大学Gansu Agricultural University
2 NTS309 强冬性Strong winterness 甘肃农业大学Gansu Agricultural University
3 GAU24 强冬性Strong winterness 甘肃农业大学Gansu Agricultural University
4 16-2444 强冬性Strong winterness 甘肃农业大学Gansu Agricultural University
5 天油14号Tianyou 14 冬性Winterness 天水市农业科学研究所Tianshui Agricultural Science Research Institute
6 天油2238 Tianyou 2238 冬性Winterness 天水市农业科学研究所Tianshui Agricultural Science Research Institute
7 天油2288 Tianyou 2288 冬性Winterness 天水市农业科学研究所Tianshui Agricultural Science Research Institute
8 甘杂9号Ganza 9 弱冬性Weak winterness 西北农林科技大学Northwest Agriculture & Forestry University
9 秦早1号Qinzao 1 弱冬性Weak winterness 陕西省杂交油菜中心Shaanxi Hybrid Rapeseed Center

Fig. 1

Cytochemical staining of leaves in different winter rapeseed (Brassica napus L.) cultivarsd under cold stress"

Table 2

Evaluation of cold tolerance in different winter rapeseed (Brassica napus L.) cultivars"

品种
Varieties
越冬率
Overwintering rate (%)
根颈直径
Collar diameter (mm)
干重
Dry weight
(g)
根/冠
Root/crown
生长点高度
The height of growth point (cm)
GAU30 75.00±3.06a 26.38±0.20ab 63.84±6.07ab 0.15±0.02ab 3.17±0.18de
NTS309 74.00±2.31a 27.40±1.49ab 67.41±8.99ab 0.15±0.03ab 3.07±0.25e
GAU24 73.67±2.33a 25.63±1.59b 71.41±2.93ab 0.14±0.03ab 3.27±0.14 cde
16-2444 74.33±2.19a 25.61±1.94b 52.65±6.68b 0.20±0.03a 3.13±0.29de
天油14号Tianyou 14 65.00±3.21b 27.01±0.85ab 80.82±4.53a 0.16±0.04ab 4.06±0.12abc
天油2238 Tianyou 2238 61.67±2.19b 23.20±2.06bc 75.17±10.94ab 0.12±0.01ab 3.96±0.06bcd
天油2288 Tianyou 2288 64.00±2.08b 19.04±0.97c 50.54±8.15b 0.11±0.01b 4.14±0.57ab
甘杂9号Ganza 9 57.00±2.53bc 25.77±0.40b 57.16±4.03ab 0.14±0.01ab 4.75±0.17ab
秦旱1号Qinzao 1 49.67±2.60c 30.65±1.96a 63.80±2.41ab 0.10±0.000b 4.89±0.25a
与越冬率的相关性
The correlation with overwintering rate
1 -0.191(P=0.622) 0.012 (P=0.976) 0.677* (P=0.045) -0.972** (P<0.001)

Fig. 2

Measurement of growth cone height in different winter rapeseed (Brassica napus L.) a1-a3: Anatomical morphology of the shoot apical meristem (SAM) in NTS309, Tianyou 14, and Qinzao 1, respectively; the distance between the two red lines indicate meristem height; b: Schematic model illustrating how plant hormones regulate SAM morphology. AUX: Auxin, GA: Gibberellic acid, CK: Cytokinin, IAA: Indole acetic acid, GA19: Gibberellic acid 19, IAA-Glu: Indole-3-acetyl-glutamic acid, ME-IAA: Methyl indole-3-acetate, IAM: 3-Indolylacetylamine, IAN: Indole-3-acetonitrile, mT9G: 3-[[(9-beta-D-xylopyranosyl-9H-purin-6-yl)amino] methyl]phenol, iP9G: Isoaminal adenine-9-glucoside, BAP9G: 6-Benzyloxy-9-(α-D-xylopyranosyl) pyrrole; c1-c3: Paraffin sections of the growth cone in NTS309, Tianyou 14, and Qinzao 1, respectively; the red arrow points to newly differentiated meristematic tissue. Bar=200 μm"

Fig. 3

Hormonomics analysis of different winter rapeseed (Brassica napus L.) cultivars a: Venn diagram of differential metabolites; b: Cluster heatmap of differential metabolites; c: KEGG enrichment analysis of NTS309 vs Tianyou 14; d: KEGG enrichment analysis of NTS309 vs Qinzao 1; e: KEGG enrichment analysis of Tianyou 14 vs Qinzao 1"

Fig. 4

Transcriptome analysis of different winter rapeseed (Brassica napus L.) cultivars a: Venn diagram of differential genes; b: Volcano plot of differential gene in NTS309 vs Tianyou 14; c: Column diagram of GO analysis in NTS309 vs Tianyou 14; d: KEGG enrichment analysis of NTS309 vs Tianyou 14; e: KEGG enrichment analysis of Tianyou 14 vs Qinzao 1"

Fig. 5

Integrated metabolomic and transcriptomic analyses of winter rapeseed with different cold resistance a: Hormonomic PCA; b: Transcriptomic PCA; c: KEGG enrichment of NTS309 vs Tianyou 14; d: KEGG enrichment of NTS309 vs Qinzao 1; e: O2PLS metabolite loading plot,IAA-Phe: Indoleacetic acid-phenylalanine, IAA-Glu: Indole-3-acetyl-glutamic acid, ME-IAA: Methyl indole-3-acetate, IAM: 3-Indolylacetylamine, IAN: Indole-3-acetonitrile, IPA: Indole-3-butyrate, iP9G: Isoaminal adenine-9-glucoside, tZR: trans-Zeatin, MeSAG: Methyl Salicylate Glucoside, JA-ILE: Jasmonic acid-isoleucine; f: O2PLS gene loading plot"

Table 3

Analysis of endogenous hormone contents in different winter cultivars of rapeseed"

缩略词Index 化合物
Compounds
类别
Class
NTS309 天油14号
Tianyou 14
秦早1号
Qinzao 1
P
P value
IAA 吲哚-3-乙酸Indole-3-acetic acid 生长素Auxin 24.37±0.56a 17.49±0.19b 28.34±3.34a 0.021
IAA-Glu 吲哚乙酸-谷氨酸Indole-3-acetyl glutamic acid 生长素Auxin 1.79±0.17a 0.38±0.04b 1.58±0.15a <0.001
MEIAA 吲哚-3-乙酸甲酯Methyl indole-3-acetate 生长Auxin 285.43±30.95a 82.37±14.75b 276.44±18.63a 0.001
IAM 3-吲哚乙酰胺3-Indole acetamide 生长素Auxin 85.23±9.41a 22.91 ±2.23c 55.20±5.32b 0.001
IAN 吲哚-3-乙腈3-Indoleacetonitrile 生长素Auxin 1796.08±69.00a 611.95±67.05b 1597.89±93.47a <0.001
ICA 吲哚-3-甲酸Indole-3-carboxylic acid 生长素Auxin 45.63±2.15a 22.72±0.56b 22.11±0.48b <0.001
ICAld 吲哚-3-甲醛Indole-3-carboxaldehyde 生长素Auxin 156.42±24.31a 59.26 ±4.07b 108.93±8.58ab 0.011
IPA 3-吲哚丙酸3-Indolepropionic acid 生长素Auxin 0.00±0.00b 0.55±0.03a 0.00±0.00b <0.001
tZR 玉米素核苷Trans-Zeatin riboside 细胞分裂素CK 2.27±0.15a 1.60±0.15b 0.40±0.03c <0.001
BAP9G 6-苄氨基-9-(Α-D-吡喃葡萄糖基)呤
N6-Benzyladenine-9-glucoside
细胞分裂素CK 0.00±0.00b 0.11±0.01a 0.11±0.01a <0.001
mT9G 3-[[(9-BETA-D-吡喃葡萄糖基-9H-嘌呤-6-基)氨基]甲基]苯酚meta-Topolin-9-glucoside 细胞分裂素CK 0.00±0.00b 0.26±0.04a 0.32±0.05a 0.002
iP9G 异戊烯腺嘌呤-9-葡糖苷N6-Isopentenyl-adenine-9-glucoside 细胞分裂素CK 0.00±0.00c 1.71±0.20b 4.40±0.34a <0.001
GA19 赤霉素19 Gibberellin A19 赤霉素GA 0.73±0.12a 0.00±0.00b 0.00±0.00b <0.001
JA 茉莉酸Jasmonic acid 茉莉酸JA 100.61±8.17a 37.79±2.67b 36.24±4.34b <0.001
OPC-4 氧化戊烯基环戊烷丁酸
3-oxo-2-(2-(Z)-Pentenyl) cyclopentane-1-butyric acid
茉莉酸JA 45.98±3.93a 12.21±0.70b 18.33±1.96b <0.001
JA-ILE 茉莉酸-异亮氨酸Jasmonoyl-L-isoleucine 茉莉酸JA 8.09±0.71a 4.56±0.05b 0.00±0.00c <0.001
MeJA 茉莉酸甲酯Methyl jasmonate 茉莉酸JA 2.84±0.08a 0.00±0.00b 0.00±0.00b <0.001
JA-Val 茉莉酸-缬氨酸N-[(-)-Jasmonoyl]-(L)-valine 茉莉酸JA 0.11±0.004a 0.00±0.00b 0.00±0.00b <0.001
ABA-GE 脱落酸葡萄糖酯ABA-glucosyl ester 脱落酸ABA 21.61±1.07a 0.00±0b 0.00±0b <0.001
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