Role of the AtERF49 in the Responses to Salt-alkali Stress in Arabidopsis

doi:10.13560/j.cnki.biotech.bull.1985.2022-0479

Abstract

Abstract:

Saline-alkali stress is one of the main environmental constraints that cause crop failures. AP2/ERF（APELATA2/ethylene response factors）transcription factors play important roles in plant growth, development and response to abiotic stress. Investigating the responses of AtERF49 to Saline-alkali stress would lay a foundation for explaining the molecular mechanism of plant AtERF49 involving in Saline-alkali stress. Arabidopsis wild-type, namely, Col-0, AtERF49-overexpressed lines of Arabidopsis and CRISPR/Cas9 mutant erf49 of Arabidopsis were treated with 150 mmol/L Saline-alkali solution（NaHCO₃ and Na₂CO₃ molar ratio 9∶1 mixed）. The RT-qPCR assays were used to analyze the basic characteristics and expression pattern of this gene under saline-alkali stress and photosynthetic response. Results showed that the leaves of erf49 mutant were wilt and bleach under Saline-alkali stress, and the leaves of AtERF49-overexpressed lines turned to be yellow slightly. Moreover, the overexpression of AtERF49 up-regulated the expression of Saline-alkali stress inducible genes（RD29A and RAB18）and photosynthetic gene rbcL under Saline-alkali stress. Chlorophyll fluorescence parameters assay of Arabidopsis leaves revealed that actual photochemical efficiency（Y(II)）and non-photochemical quenching coefficient（qP）in AtERF49-overexpressed plants were significantly higher than that on Col-0, and the photodamage（NO）and photochemical quenching coefficient（qN）were significantly lower compared to Col-0. However, the opposite trend was apparent in erf49 mutant. These results suggested that AtERF49 might participate in response to Saline-alkali stress through regulating its downstream Saline-alkali stress-responsive genes or affecting photosynthesis efficiency.

Key words: AtERF49, salt-alkali stress, photosynthesis, Arabidopsis

RUAN Hang, DUO Hao-yuan, FAN Wen-yan, LV Qing-han, JIANG Shu-jun, ZHU Sheng-wei. Role of the AtERF49 in the Responses to Salt-alkali Stress in Arabidopsis[J]. Biotechnology Bulletin, 2023, 39(1): 150-156.

Figures/Tables 6

Table 1 Primers used for fluorescence quantitative PCR

引物名称Primer name	引物序列Primer sequence（5'-3'）
RAB18-F	TCCAGCTCTAGCTCGGAGGATGA
RAB18-R	GGATCCCATGCCGCCCATCG
RD29A-F	GCCGGAATCTGACGGCGGTT
RD29A-R	CCCGTCGGCACATCCTTGTCG
AtUBC30-F	TCACTTCCCACCAGATTACCC
AtUBC30-R	TCGACAGAAGAACCTTGGATACG
AtERF49-F	AGACTACACCAAGCAGCAACACC
AtERF49-R	TTGGATGAACACGGCGACTCAG
rbcl-F	TTGGCAGCATTCCGAGTAACTCCT
rbcl-R	CTGGTAAGTCCATCGGTCC
psaA-F	CTACTTTGCCACCCACTGC
psaA-R	TGAGTGCTTTAGGGCGTCC

Fig. 1 Analysis of AtERF49 expression under Saline-alkali stress *: P<0.05; ***: P<0.001. The same below

Fig. 2 Overexpression of AtERF49 gene increasing Arabidopsis resistance to Saline-alkali stress A: Phenotypic analysis of AtERF49 transgenic Arabidopsis phenotype with Saline-alkali stress, the scale is 2 cm. B: Relative expression of AtERF49 in wild type and transgenic lines

Fig. 3 Expression analysis of RAB18（A）and RD29A（B）in AtERF49 transgenic lines

Fig. 4 Effects of Saline-alkali stress on the chlorophyll fluorescence parameters of leaves of AtERF49 transgenic Arabidopsis A: Energy dissipation quantum yield. B: Actual quantum yield of photosystem II. C: Coefficient of non-photochemical quenching. D: Coefficient of photochemical quenching

Fig. 5 Relative expressions of rbcL and psaA in AtERF49 transgenic lines

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