The Inositol-Phosphate Synthase Gene，MdMIPS1，Enhances Heat Stress Tolerance in Transgenic Apple Plants

doi:10.16420/j.issn.0513-353x.2024-0997

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (12): 3143-3156.doi: 10.16420/j.issn.0513-353x.2024-0997

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Next Articles

The Inositol-Phosphate Synthase Gene，MdMIPS1，Enhances Heat Stress Tolerance in Transgenic Apple Plants

SUN Wenjing, LI Xiaoyan, ZHANG Jingyun, ZHOU Hui, HUANG Xiaojun, MA Fengwang^*(), GONG Xiaoqing^*()

State Key Laboratory for Crop Stress Resistance and High-Efficiency Production，Shaanxi Key Laboratory of Apple，College of Horticulture，Northwest A & F University，Yangling，Shaanxi 712100，China

Received:2025-08-08 Revised:2025-10-20 Online:2025-12-25 Published:2025-12-20
Contact: MA Fengwang, GONG Xiaoqing

Abstract

Abstract:

In order to reveal the role of Myo-inositol（MI）in apple plants responding to high-temperature stress，apple plants were first exogenously supplied with MI and treated with high temperature stress. The results showed that high-temperature stress significantly caused leaf yellow and wilting，increased the contents of reactive oxygen species（ROS），malondialdehyde（MDA），and relative electrolyte leakage（REL），decreased the contents of the chlorophyll in apple plants. However，100 µmol · L^-1exogenous MI treatment significantly alleviated the damage caused by high temperature. In addition，MdMIPS1 encoding the key rate-limiting enzyme in the MI biosynthesis，expressed highly under high-temperature stress in apple plants. The transgenic lines overexpressed MdMIPS1（OE，high MI content）were obtained in our previous study. They had higher endogenous content of MI，compared with wild type（WT）plants. And these OE lines were also used in this work and treated with high temperature stress. Compared with WT plants under high temperature stress，the OE lines exhibited slighter chlorisis and wilting in leaves. The contents of REL were lower and the contents of the chlorophyll were higher in OE plants than in WT. In addition，the stomatal aperture were larger in OE leaves，and the photosynthetic efficiency were stronger in OE plants than in WT. Meanwhile，the OE plants possessed less ROS accumulation and higher antioxidant enzyme activity than WT. The expression levels of several genes encoding the heat shock protein were also higher in OE plants than in WT under high temperature stress. In sum，all these results indicated that increasing the MI levels can significantly enhance tolerance to high temperature stress in apple plants.

Key words: apple, Myo-inositol, MdMIPS1, high-temperature stress

SUN Wenjing, LI Xiaoyan, ZHANG Jingyun, ZHOU Hui, HUANG Xiaojun, MA Fengwang, GONG Xiaoqing. The Inositol-Phosphate Synthase Gene，MdMIPS1，Enhances Heat Stress Tolerance in Transgenic Apple Plants[J]. Acta Horticulturae Sinica, 2025, 52(12): 3143-3156.

Figures/Tables 8

Table 1 Primer sequences of qRT-PCR

引物名称Primer name	正向引物序列（5′-3′）Forward primer sequence	反向引物序列Reverse primer sequence
qEF-1α	ATTCAAGTATGCCTGGGTGC	CAGTCAGCCTGTGATGTTCC
qMdMIPS1	AGGCCAAGTTCCACTCATTCCAC	CAAGCCCTCAGTATGTTCTCCAG
qMdHY5	AACTAGCATCCAAAGCTAGCC	CCACAGTCTGCAGTTTCACAAG
qMdDREB2A	CAGAGAACTTCACAACCGAC	CCAGTGCTCATGGCAAGGTTTT
qMdHSP17.3	AAGTTTGTGTTGCCGGAG	ATGTTATAATCCCTGCACGA
qMdHSP70-2	CTCTGTCTCTCGGTTTGGAG	GTCTTGTCCTCGGCAGAAA
qMdHSP70-1	CAAGCTGAAAGAGCTCAAGG	CTGTCCCAGCTGCATTACTT
qMdHSP90-5	ACACTCACTTCACCACTGAG	CATTTGAATCCACCACACCC

Fig. 1 Effect of exogenous MI on heat resistance of apple under high temperature stress A：The performance；B：MDA content；C：Total chlorophyll content；D：Electrolyte leakage. The data were shown as the mean of three replicates，and univariate analysis of variance and duncan multiple comparison were used，with different letters indicating significant differences between the data（P < 0.05）. The same below

Fig. 2 Effect of exogenous MI on antioxidant capacity of apple under high temperature stress

Fig. 3 Analysis of expression of apple MdMIPS1 A：Relative expression levels in tissues；B：Spatio-temporal expression patterns in leaves

Fig. 4 Analysis of heat resistance of MdMIPS1 transgenic apple plants under high temperature stress A：The performance of plant and leaf；B：Electrolyte leakage；C：Chlorophyll content. WT：GL-3 wild-type. OE-4/OE-6：Transgenic apple plants with overexpression of MdMIPS1. The same below

Fig. 5 Analysis of photosynthetic capacity of MdMIPS1 transgenic apple plants under high temperature stress A：The chlorophyll fluorescence；B：Fv/Fm；C：Tr；D：Pn；E：Ci；F：Leaf stomata；G：Stomatal opening；H：Gs. Different letters indicated WT，OE-4 and OE-6 strains have significant differences in control or heat stress data significant differences between the data（P < 0.05）

Fig. 6 Analysis of antioxidant capacity of MdMIPS1 transgenic apple plants under high temperature stress A：NBT and DAB staining；B： $\mathrm{O}_{2}^{\overline{·}}$ content；C：H2O2 content；D：SOD activity；E：POD activity

Fig. 7 Relative expression levels of heat shock proteins genes in WT and OE plants under high temperature stress Row Z-score normalization

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The Inositol-Phosphate Synthase Gene，MdMIPS1，Enhances Heat Stress Tolerance in Transgenic Apple Plants

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