The role of a cytokinin signaling pathway type-B ARR transcription factor， LpARR10， in cadmium tolerance of perennial ryegrass

doi:10.11686/cyxb2021097

Abstract

Abstract:

Cadmium （Cd） is one of the major elements causing heavy metal pollution in mining areas that can severely inhibit plant growth and development. It is known that cytokinin （CTK） can enhance plant Cd tolerance， but its underlying mechanism is still unclear. In this study， a CTK signal pathway gene， LpARR10， was cloned in perennial ryegrass that has six exons and encodes 598 amino acids. Evolutionary analysis indicated that LpARR10was clustered together with the Arabidopsis B-type ARR transcription factors （B-ARRs） and closely related to AtARR10 and AtARR12. Protein sequence analysis demonstrated that LpARR10 has typical functional domains of B-ARRs featured with a phosphate receptor domain， phosphorylation sites， and metal ion binding sites. Expression analysis showed that both CTK （25 μmol·L^-1 6-BA） and Cd （200 μmol·L^-1 CdCl₂） significantly promoted the transcription of LpARR10 in roots of perennial ryegrass after 0.5， 2， 6， 12， 24 and 48 h of treatment and in leaves after 2 and 6 h. When grown in MS medium， there was no significant difference in root length and fresh weight between transgenic lines and wild type plants. However， the root length and fresh weight of the transgenic lines were significantly higher than those of the wild type plants when grown in MS medium with Cd （180 μmol·L^-1 CdCl₂）. In conclusion， LpARR10 plays an important role in cytokinin-regulated plant Cd tolerance.

Key words: perennial ryegrass, cytokinin, LpARR10 gene, cadmium stress, transgenic Arabidopsis

Qing ZHANG, Jing XING, Jia-ming YAO, Ting-chao YIN, Xin-ru HUANG, Yue HE, Jing ZHANG, Bin XU. The role of a cytokinin signaling pathway type-B ARR transcription factor， LpARR10， in cadmium tolerance of perennial ryegrass[J]. Acta Prataculturae Sinica, 2022, 31(5): 135-143.

Figures/Tables 7

Table 1 Primers used in the study

引物名称Primer	上游引物序列Forward primer sequence （5′-3′）	下游引物序列Reverse primer sequence （5′-3′）
LpARR10-CDS	ATTTggatccATGCGCGTGCTCGCCGTC	ATTTactagtGACCAGCTGCCAGTCCCCAT
LpARR10-qRT	GATGTCGGGAATACGGGCGG	GAGCCCCCTTTGCAGCCTAC
AtTUBULIN4-qRT	CTGTTTCCGTACCCTCAAGC	AGGGAAACGAAGACAGCAAG
Bar gene	TCAAATCTCGGTGACGGGCAGGAC	AACCGCAGGAGTGGACGGACGA

Fig.1 Phylogenetic tree of LpARR10and Arabidopsis ARRtranscription factors

Fig.2 Genetic structure， amino acid sequence and function domain analysis of LpARR10 and its homologus genes

Fig.3 Expression analysis of LpARR10 gene in perennial ryegrass leaves and roots treated with cytokinin

Fig.4 LpARR10 gene expression in leaves and roots of perennial ryegrass under cadmium treatment

Fig.5 Identification of LpARR10 overexpression transgenic lines

Fig.6 Fresh weight and root length in transgenic lines and wild type Arabidopsis when treated with Cd

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