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Acta Agronomica Sinica ›› 2025, Vol. 51 ›› Issue (1): 58-67.doi: 10.3724/SP.J.1006.2025.44092

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

Preliminary investigation of the SiLTP1: a lipid transfer protein gene involved in the salt tolerance of foxtail millet

MENG Fan-Hua1, LIU Min1(), SHEN Ao1,2, LIU Wei1,*()   

  1. 1Institute of Crop Germplasm Resources, Shandong Academy of Agricultural Sciences, Jinan 250100, Shandong, China
    2School of Ecological Technology & Engineering, Shanghai Institute of Technology, Shanghai 201418, China
  • Received:2024-06-06 Accepted:2024-09-18 Online:2025-01-12 Published:2024-10-10
  • Contact: *E-mail: wheiliu@163.com
  • About author:**Contributed equally to this work
  • Supported by:
    Agricultural Fine Seed Project of Shandong Province(2021LZGC006);Key Research and Development Project of Shandong Province(2021LZGC025);National Natural Science Foundation of China(32201736);National Natural Science Foundation of China(32171955);Agricultural Science and Technology Innovation Project of Shandong Academy of Agricultural Sciences(CXGC2023F13)

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Abstract:

Soil salinization poses a significant barrier to agricultural productivity. Enhancing the salt tolerance of crops is crucial for the effective utilization of saline-alkali land, thereby ensuring food security and improving agricultural efficiency. Lipid transfer proteins (LTPs) are a class of small, widely distributed proteins in higher plants, playing crucial roles in plant growth, signal transduction, and responses to various biotic and abiotic stresses. In a previous study, a lipid transfer protein gene, designated SiLTP1, was cloned from foxtail millet. In this study, we constructed both a prokaryotic expression vector and a plant binary overexpression vector for SiLTP1. The prokaryotically expressed protein and four homozygous transgenic lines were successfully obtained and characterized. In vitro salt tolerance assays revealed that the SiLTP1 protein exhibited a notable degree of salt tolerance. Physiological measurements of the transgenic seedlings indicated reduced oxidative damage, as evidenced by lower malondialdehyde (MDA) accumulation, increased superoxide dismutase (SOD) activity, and reduced hydrogen peroxide (H2O2) levels under salt stress, demonstrating enhanced salt tolerance. These findings suggest that SiLTP1 plays a positive role in regulating salt tolerance in foxtail millet, potentially by mitigating oxidative stress. This study provides a theoretical foundation and valuable genetic resources for the future development and breeding of salt-tolerant foxtail millet varieties.

Key words: lipid transfer proteins, SiLTP1, salt stress, foxtail millet, salt tolerance response

Fig. 1

Prokaryotic protein expression of SiLTP1 A: PCR verification; M: DNA marker; 1-6: positive bacterial solution of pET32a-SiLTP1. B: SDS-PAGE electrophoresis; M: protein maker; 1-4: total protein expressed by pET32a-SiLTP1, induced by 0.05 mmol L-1, 0.20 mmol L-1, 0.50 mmol L-1, and 1.00 mmol L-1 IPTG for 5 h; 5-6: total protein expressed by pET32a; 7: total protein of plants without vector transformation. 32 kD band: fusion protein expressed by pET32a-SiLTP1; 21 kD band: the tagged protein expressed by the vector."

Fig. 2

Detection of salt tolerance of pET32a-SiLTP1 strain with spot method Phenotypes of pET32a-SiLTP1 and pET32a on LB with 0 mmol L-1, 100 mmol L-1, 200 mmol L-1, 300 mmol L-1, 500 mmol L-1, and 700 mmol L-1 NaCl."

Fig. 3

Growth curves of pET32a-SiLTP1 (blue) and pET32a (orange) in LB liquid medium with 200 mmol L-1 NaCl * indicates a significant difference at the 0.05 probability level."

Fig. 4

Identification and screening of SiLTP1 overexpressing lines A: hygromycin screening; B: PCR identification of overexpressing plants; M: DNA marker; 1-4: overexpressing lines of T3 generation; 5: Ci846. C: relative expression levels of SiLTP1 in Ci846 and overexpression lines (OE1-OE4). Different lowercase letters indicate significant differences at the 0.05 probability level."

Fig. 5

Phenotypes of control and SiLTP1 overexpressing lines before and after salt treatment A: phenotypes of control and SiLTP1 overexpressing plants before and after 100 mmol L-1 NaCl treatment for seven days; B: analyze of root and shoot length before and after 100 mmol L-1 NaCl treatment for seven days. Different lowercase letters indicate significant differences at the 0.05 probability level."

Fig. 6

Test of physiological indexes and DAB staining of Ci846 and SiLTP1 overexpressing lines before and after salt treatment Different lowercase letters indicate significant differences at the 0.05 probability level."

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