Endophytic fungal GG22 protein improves growth of Carthamus tinctorius L. seedling under stress conditions

doi:10.3969/j.issn.1004-1524.20240691

Acta Agriculturae Zhejiangensis ›› 2025, Vol. 37 ›› Issue (6): 1193-1202.DOI: 10.3969/j.issn.1004-1524.20240691

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Endophytic fungal GG22 protein improves growth of Carthamus tinctorius L. seedling under stress conditions

ZHOU Danning¹^,²(), XU Jiao¹, BAI Jing³, LIU Xiangnan¹, ZHU Yunhao¹^,²^,^*()

1. School of Pharmacy, Henan University of Chinese Medicine, Zhengzhou 450046, China
2. Respiratory Disease Diagnosis and Treatment and New Drug of Henan Collaborative Innovation Center, Zhengzhou 450046, China
3. Nanyang Vocational College, Nanyang 474550, Henan, China

Received:2024-07-31 Online:2025-06-25 Published:2025-07-08

Abstract

Abstract:

To investigate the effects of the protein from endophytic fungus Fusarium oxysporum GG22 in Rehmannia glutinosa(GG22 protein) on plant growth, different fractions of GG22 protein (designated as G20, G40, G60, and G80) were obtained using graded precipitation with ammonium sulphate. Safflower(Carthamus tinctorius L.) seedlings were treated with these GG22 protein fractions (G20, G40, G60 and G80) to preliminarily explore their impact on seedling growth under three stress conditions(high temperature, low temperature, and salinity). The relative expression levels of genes involved in plant immunity—ARD1 (activated disease resistance gene 1), EDS1 (enhanced disease susceptibility gene 1), PAD4 (plant antitoxin-deficient gene 4), NPR(non-expressors of pathogenesis related gene), and CDKs(cyclin-dependent kinase gene), under stress conditions were analyzed using quantitative real-time PCR (qRT-PCR). The results showed that the G60 and G80 groups significantly influenced safflower seedling growth under various stress conditions. Under high-temperature stress, the fresh weight of seedlings in the G80 group was 1.41 times that of the control group, while the G60 group was 1.39 times that of the control group. Under low-temperature stress (G80 group) and salt stress (G60 group), shoot length, fresh weight, and dry weight of seedlings significantly increased compared with the control. GG22 protein treatment alleviated stress-induced damage to seedling roots, with the G60 and G80 groups showing significant effects. Additionally, the expression of ARD1, EDS1, PAD4, NPR, and CDKs genes were upregulated to varying degrees in the G60 and G80 groups under all three stress conditions. Proteomic analysis identified three protein sequences with high similarity to known elicitor proteins—Aspf2, PEMG1 and PeaT1, which were selected as candidate elicitors for subsequent isolation, purification, and functional characterization. In conclusion, the protein from Fusarium oxysporum GG22 can improve safflower seedling growth under stress conditions and may contain elicitor proteins that activate plant immune responses, suggesting its potential as a plant immune elicitor.

Key words: endophytic fungi, Fusarium oxysporum, stress, activator protein, Carthamus tinctorius L.

CLC Number:

S567.21

ZHOU Danning, XU Jiao, BAI Jing, LIU Xiangnan, ZHU Yunhao. Endophytic fungal GG22 protein improves growth of Carthamus tinctorius L. seedling under stress conditions[J]. Acta Agriculturae Zhejiangensis, 2025, 37(6): 1193-1202.

Figures/Tables 6

Table 1 Primer information

名称 Name	正向引物(5'→3') Forward primer(5'→3')	反向引物(5'→3') Reverse primer(5'→3')	扩增长度 Amplification length/bp
CtCDKs	ATGGCTGCCTCTCTTTCCTC	ATGTTGAAAGGTTGGGGCAG	249
CtNPR	TCCCGCCAAAATCTTGAAAT	ATGCACCTATGAATGCCCAC	137
CtARD1	ATCTCAGCCTTCGCCCACCT	GGTCGGATTTGTCGGATGTG	133
CtEDS1	GAAGGTGCCAGTTCCGAGAC	AGCAAGATTCCGACCTGTGT	213
CtPAD4	CGCTATGGCTTCTCTCCTATC	GAGCGAACAACAGCCGTG	182
Ct60S	GGCAACACTGGTCTTGATCCAT	CTGCGATTGTTGGATAATGGATG	92

Fig.1 Effects of various fractions of GG22 protein on growth of safflower seedlings under stress conditions CK indicates blank control, G20, G40, G60 and G80 indicate treatments with the corresponding proteins. * represents P<0.05, ** represents P<0.01, *** represents P<0.001, **** represents P<0.000 1.

Table 2 Effects of various fractions of GG22 protein on root activity of safflower seedlings under stress conditions μg·g-1·h-1

处理 Treatment	根系活力Root activity
处理 Treatment	正常处理Normal	高温胁迫Heat-temperature stress	低温胁迫Low-temperature stress	盐胁迫Salt stress
CK	53.83±2.348 c	41.93±1.24 b	37.31±7.32 d	47.39±1.71 b
G20	59.91±3.061 b	37.85±1.52 b	35.77±1.73 d	49.51±4.37 b
G40	54.17±2.919 c	39.94±1.35 b	48.14±1.48 c	50.57±0.64 b
G60	62.45±0.859 ab	69.87±3.27 a	59.07±0.53 b	53.63±0.36 ab
G80	66.10±0.703 a	42.42±3.64 b	82.59±1.04 a	58.32±0.85 a

Fig.2 Effects of various fractions of GG22 protein on the expression of immunity-related genes of safflower seedlings under stress conditions A1, A2, A3, and A4 indicate G20, G40, G60 and G80 treatments under high temperature stress, respectively; B1, B2, B3 and B4 indicate G20, G40, G60 and G80 treatments under low-temperature stress, respectively; C1, C2, C3 and C4 indicate the treatment with G20, G40, G60, and G80 under salt stress, respectively.

Fig.3 Electrophoresis results of different GG22 protein fractions

Fig.4 Qualitative identification of proteomes A, Distribution of apparent relative molecular weight; B, Venn diagram for identification of different fractions of proteins.

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Endophytic fungal GG22 protein improves growth of Carthamus tinctorius L. seedling under stress conditions

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