Overexpression of TPP from Larix gmelinii Enhanced Salt Tolerance of the Transgenic Arabidopsis thaliana

doi:10.13523/j.cb.2203056

China Biotechnology

2022, Vol. 42

Issue (8): 21-29 DOI: 10.13523/j.cb.2203056

Overexpression of TPP from Larix gmelinii Enhanced Salt Tolerance of the Transgenic Arabidopsis thaliana

ZHANG Yan-xia¹,ZHANG Xu-ting¹,JIA Yong-hong²,LEI Feng-yan³,WANG Jing¹,WANG Rui-gang^1,⁴,LI Guo-jing^1,^**(

)

1. Inner Mongolia Key Laboratory of Plant Stress Physiology and Molecular Biology, College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China
2. Vocational and Technical College, Inner Mongolia Agricultural University, Baotou 014109, China
3. Agriculture and Animal Husbandry Bureau of Ongniud Banner of Tongliao City, Tongliao 028000, China
4. Inner Mongolia Enterprise Key Laboratory of Tree Breeding, Mengshu Ecological Construction Group Co.,Ltd., Hohhot 011517, China

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Abstract

Larix gmelinii is a very important coniferous tree species for afforestation. It has the characteristics of rapid growth in early stage, strong stress resistance and good ecological benefits. Trehalose participates in the regulation of drought, cold, salt damage and other stresses. Trehalose-6-phosphate phosphatase (TPP) is an important enzyme in the trehalose synthesis pathway. In this study, the full-length sequence of LgTPPI.1 was screened from the transcriptome of L. gmelinii under stress and its coding sequence(CDS) was cloned. The recombinant vector was constructed and homozygous lines of transgenic Arabidopsis thaliana overexpressing LgTPPI.1 were obtained. The results showed that the full-length CDS of LgTPPI.1 was 1 236 bp, encoding 411 amino acids; the expression level of LgTPPI.1 was lower in roots and stems, but higher in needles. Under salt treatment, the LgTPPI.1 overexpression lines conferred stronger tolerance than the wild type A. thaliana, with elevated trehalose and proline content; increased superoxide dismutase(SOD) and catalase(CAT) activity and up-regulated expresssion of the stress-responsive marker genes. These results indicated that gymnosperm utilized similar trehalose pathway as angiosperm to telerate abiotic stress. This study provided a theoretical basis for further analysis of the function of trehalose synthetic genes and the response mechanism of conifers under stress.

Key words： Larix gmelinii Salt stresses Trehalose TPP Arabidopsis thaliana

Received: 28 March 2022 Published: 07 September 2022

ZTFLH:

Q812

Corresponding Authors: Guo-jing LI E-mail: liguojing@imau.edu.cn

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	Yan-xia ZHANG
	Xu-ting ZHANG
	Yong-hong JIA
	Feng-yan LEI
	Jing WANG
	Rui-gang WANG
	Guo-jing LI

Cite this article:

ZHANG Yan-xia,ZHANG Xu-ting,JIA Yong-hong,LEI Feng-yan,WANG Jing,WANG Rui-gang,LI Guo-jing. Overexpression of TPP from Larix gmelinii Enhanced Salt Tolerance of the Transgenic Arabidopsis thaliana. China Biotechnology, 2022, 42(8): 21-29.

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https://manu60.magtech.com.cn/biotech/10.13523/j.cb.2203056 OR https://manu60.magtech.com.cn/biotech/Y2022/V42/I8/21

Table 1 The list of primers used in this study

Fig.1 Electrophoresis gel of the cloning products of LgTPPI.1 and the digestion verification of the recombinant vector (a)PCR products of LgTPPI.1 CDS (b)35S∷HA-LgTPPI.1 recombinant vector digestion and identification M1: DL2000 marker; M2: DL10000 marker; Lanes 1 and 2: PCR product of LgTPPI.1 CDS; Lanes 3: 35S∷HA-LgTPPI.1 recombinant vector digestion with Sal I and Sac I;Lane 4: Plasmid control

Fig.2 The gene and its encoded protein sequence of LgTPPI.1

Fig.3 Phylogenetic analysis of LgTPPI.1 and its homologous proteins from A. thaliana

Fig.4 The expression pattern analysis of LgTPPI.1 in different tissues of L. gmelinii

Fig.5 Detection of LgTPPI.1 expression level of the homozygous lines expressing 35S∷HA-LgTPPI.1 in A. thaliana

Fig.6 Comparison of tolerance to salt stress between the LgTPPI.1 overexpressing lines and wild type A. thaliana

Fig.7 Physiological indicators assay of LgTPPI.1 overexpressing lines and wild type A. thaliana under salt stress

Fig.8 Detection of stress responsive marker genes of the LgTPPI.1 overexpressing lines and wild type A. thaliana

Fig.9 Comparison of the trehalose content of the homozygous lines overexpressing LgTPPI.1 and wild type A. thaliana before and after salt stress


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