Scientia Agricultura Sinica ›› 2023, Vol. 56 ›› Issue (1): 193-202.doi: 10.3864/j.issn.0578-1752.2023.01.015

• RESEARCH NOTES • Previous Articles    

Functional Analysis of MiZAT10A and MiZAT10B Genes in Mango

MO WenJing(),ZHU JiaWei(),HE XinHua,YU HaiXia,JIANG HaiLing,QIN LiuFei,ZHANG YiLi,LI YuZe,LUO Cong()   

  1. College of Agriculture, Guangxi University/State Key Laboratory of Subtropical Agricultural Biological Resources Protection and Utilization/National Experimental Teaching Demonstration Center of Plant Science, Nanning 530004
  • Received:2022-03-02 Accepted:2022-05-12 Online:2023-01-01 Published:2023-01-17
  • Contact: Cong LUO E-mail:tuanzy97616@163.com;zhujiaweiii1206@163.com;22003luocong@163.com

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Abstract:

【Objective】Zinc finger protein (ZFP) plays an important role in plant abiotic stress response. Therefore, to provide a theoretical basis for stress resistance breeding, this study aimed to analyze the response of two zinc finger protein genes of MiZAT10A and MiZAT10B transgenic Arabidopsis to abiotic stresses, such as salt, drought, heavy metals and exogenous hormones. 【Method】 The promoter cis acting elements and motif of mango MiZAT10A and MiZAT10B genes were predicted and analyzed by online software PLACE and MEME, respectively. The chromosome location map was drawn by TBtools software and SiJiMi gene annotation file (GFF file and unpublished). Tissue expression patterns of MiZAT10A and MiZAT10B genes were analyzed by qRT-PCR. The overexpression vectors of MiZAT10A and MiZAT10B genes were constructed and transformed into Arabidopsis thaliana by Agrobacterium floral-dip method. The phenotype of MiZAT10A and MiZAT10B transgenic plant were observed and recorded under salt, drought, heavy metals, abscisic acid and gibberellin treatments. 【Result】 Promoter cis element analysis showed that there were many light response elements, hormone response elements and abiotic stress response elements in the promoter region of MiZAT10A and MiZAT10B genes. Expression analysis showed that MiZAT10A and MiZAT10B were highly expressed in buds and flowers. 9 of MiZAT10A and 14 of MiZAT10B transgenic Arabidopsis strains were obtained. Overexpression of MiZAT10A and MiZAT10B significantly resulted early flowering compared with the control lines. The root length of MiZAT10A and MiZAT10B overexpressing transgenic Arabidopsis was significantly longer than that of control lines under salt stress, drought stress, heavy metal stress, GA3 and ABA hormone treatments. 【Conclusion】 Overexpression of MiZAT10A and MiZAT10B not only promoted transgenic Arabidopsis flowering early but also improved salt, drought, heavy metals and exogenous hormones GA3 and ABA resistance.

Key words: mango, abiotic stress, zinc finger protein, expression, function analysis

Table 1

Primer sequences used in this study"

引物 Primer 序列 Sequences (5′-3′) 用途 Usage
AtActin2-F GCAGAGCGGGAAATTGTAAG 半定量Semi-quantitative
AtActin2-R GTACAGATCCTTCCTGATATCC 半定量Semi-quantitative
MiActin-F CCGAGACATGAAGGAGAAGC 实时荧光定量qRT-PCR
MiActin-R GTGGTCTCATGGATACCAGCA 实时荧光定量qRT-PCR
MiZAT10A-F GCTAAGCGCAAAAGGTCGAA 实时荧光定量qRT-PCR
MiZAT10A-R GGTCGTAGCAGCTGATGGAG 实时荧光定量qRT-PCR
MiZAT10B-F GCGТССТСАCAACCATCCAA 实时荧光定量qRT-PCR
MiZAT10B-R GGAGGTTGACTGCTCGTCG 实时荧光定量qRT-PCR

Fig. 1

Sequence analysis of MiZAT10A and MiZAT10B genes A: Chromosomal mapping of MiZAT10A and MiZAT10B genes; B: Conserved motif distribution of MiZAT10A and MiZAT10B proteins"

Fig. 2

Promoter cis-regulatory element analysis of MiZAT10A and MiZAT10B genes"

Fig. 3

Tissue expression analysis of MiZAT10A and MiZAT10B genes A: MiZAT10A tissue expression; B: MiZAT10B tissue expression. Different lowercase letters represent significant difference at P<0.05 level. The same as below"

Fig. 4

Flowering phenotypic analysis of overexpressed MiZAT10A and MiZAT10B genes A1: Flowering photos of overexpressed MiZAT10A gene in Arabidopsis thaliana; A2: Flowering data of overexpressed MiZAT10A gene in Arabidopsis thaliana; B1: Flowering photos of overexpressed MiZAT10B gene in Arabidopsis thaliana; B2: Flowering data of overexpressed MiZAT10B gene in Arabidopsis thaliana"

Fig. 5

Abiotic stress resistance analysis of overexpression MiZAT10A and MiZAT10B in Arabidopsis thaliana A1: Root length data of overexpressed MiZAT10A transgenic Arabidopsis thaliana under stress; A2: Root length data of over-expressed MiZAT10B transgenic Arabidopsis under stress; B1: Root-length data of over-expressed MiZAT10A transgenic Arabidopsis under stress; B2: Root-length photos of overexpressed MiZAT10B gene in Arabidopsis; *represent significant difference at 0.05 level"

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