Cloning and salt-tolerance analysis of MsWRKY11 in alfalfa

doi:10.11686/cyxb2020426

Abstract

Abstract:

MsWRKY11 was cloned from the cDNA of Medicago sativa， andtransgenic lines of over expressed MsWRKY11 （MsWRKY11-OE） in alfalfawere obtained by transplanting PHB-MsWRKY11-Flag expression vector into alfalfa. The effect of MsWRKY11 on alfalfa tolerance to salt stress was studied by treating transgenic lines and wild type （WT） with 0 or 250 mmol·L^-1 NaCl. The results showed that MsWRKY11 was closely related to Medicago truncatula WRKY11， and the expression of MsWRKY11 was upregulated by addition of NaCl， salicylic acid and jasmonic acid. The growth rates （aboveground biomass and height）， peroxidase activity， catalase activity and ratio of K⁺/Na⁺ were increased in MsWRKY11-OE lines compared with WT， meanwhile the Na⁺ content， relative conductance， malondialdehyde content and superoxide anion content were decreased in MsWRKY11-OE lines under salt stress. The expression of salt-tolerance related genes， including MsNHX1， MsSOS3， MsAPX， MsGRX， MsNAC and MsP5CS were upregulated under salt stress. Also， the expression of those genes was higher in MsWRKY11-OE lines than in the WT under salt stress. These results indicate that MsWRKY11 was involved in the maintenance of K⁺/Na⁺ homeostatic status and stimulation of peroxidase activity under salt stress， and through these mechanisms enhanced the salt-tolerance of alfalfa.

Key words: alfalfa （Medicago sativa）, WRKY, salt-tolerance, transcription factor

Ru-yue WANG, Wu-wu WEN, En-hua ZHAO, Peng ZHOU, Yuan AN. Cloning and salt-tolerance analysis of MsWRKY11 in alfalfa[J]. Acta Prataculturae Sinica, 2021, 30(11): 157-169.

Figures/Tables 11

Table 1 qRT-PCR primer sequences

耐盐性相关基因 Salt-related gene	正向引物（5′-3′） Forward primer （5′-3′）	反向引物（3′-5′） Reverse primer （3′-5′）
EF-α	GCACCAGTGCTCGATTGC	TCGCCTGTCAATCTTGGTAACAA
MsWRKY11	CTTATAGCATCTCTACCACTAC	CTTGCCAGAGGAAATGATAG
MsNHX1	GCCTTCGTGCTTTACTATCAAC	GATTACCATTGCGTTCACTTGG
MsSOS3	GTTCTTGCTTCTGAAACACC	CCAAGAGATCGAACAAATTC
MsAPX	TCGGAACCATCAAGCACCAAGC	CAACAGCAACAACACCAGCCAAC
MsGRX	GCTGCCCAACTGTCCACCAAC	TGACTTGCCATGACTCTTTCC
MsNAC	GAAAGACTGGGATAGCGAAGAG	CTAGGTAGAATGAGAGCTGGTG
MsP5CS	CCTCGGTCGACAAAGGCTTA	CCCCCTCTTCCAACCCTAGA

Fig.1 The cloning and sequence alignment of MsWRKY11

Fig.2 Analysis of MsWRKY11 evolutionary tree and sequence alignment

Fig.3 MsWRKY11 protein structure prediction

Fig.4 The expression pattern of MsWRKY11 in leaves and roots under NaCl， SA and JA treatment

Fig.5 Identification of transgenic alfalfa

Fig.6 Phenotype analysis of MsWRKY11 transgenic lines under salt stress

Fig.7 Effect of overexpression of MsWRKY11 on membrane lipid peroxidation of alfalfa under salt stress

Fig.8 Effects of overexpression of MsWRKY11 on alfalfa antioxidant ability under salt stress

Fig.9 Content of K+， Na+ and ratio of K+/Na+ in alfalfa seedlings under salt stress

Fig.10 Expression patterns of key genes related to salt stress in transgenic alfalfa under salt stress

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