Functional Analysis of the AmNAC3 Transcription Factor Gene from <i>Ammopiptanthus mongolicus</i> in Drought and Cold Resistances

doi:10.13523/j.cb.20190805

China Biotechnology

2019, Vol. 39

Issue (8): 32-39 DOI: 10.13523/j.cb.20190805

Functional Analysis of the AmNAC3 Transcription Factor Gene from Ammopiptanthus mongolicus in Drought and Cold Resistances

REN Mei-yan,TANG Kuan-gang,XUE Min,GUO Hui-qin,WANG Mao-yan(

)

College of Life Sciences, Inner Mongolia Agricultural University, Hohhot 010018, China

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Abstract

Ammopiptanthus mongolicus is a plant with very strong tolerance to abiotic stresses.To investigate whether AmNAC3,a transcription factor gene from this species,plays roles in tolerating cold and drought, first performed the expression analysis of this gene using semi-quantitative RT-PCR.The results showed that in the seedlings cultivated in normal growth conditions, AmNAC3 had a clear basic expression and the expression level was obviously up-regulated under the drought treatment.In contrast,the expression level of AmNAC3 was weakly up-regulated under the cold stress. Then acquired the 5'-terminal sequence of AmNAC3 by 5'RACE and its full-length cDNA sequence.The whole coding region (846bp) cDNA was cloned by RT-PCR and was inserted into plant expression vector.Transgenic Arabidopsis plants of AmNAC3 was obtained by Agrobacterium mediated genetic transformation.Further analyses showed that the transgenic and wild type plants presented similar drought and cold resistance.However, the water loss rate and stomatal aperture of the detached leaves of transgenic lines were greater than those of wild type.Moreover,the transcript levels of ABI1 and ABI2, two genes related to stomatal closure,were down-regulated in transgenic seedlings.These results indicate that AmNAC3 may primarily function in the response to drought stress and regulation of stomatal closure and leaf water retention but play less role in coping with low temperature stress.

Key words： Ammopiptanthus mongolicus NAC Water loss of leaves Stomatal aperture

Received: 29 January 2019 Published: 18 September 2019

ZTFLH:

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Corresponding Authors: Mao-yan WANG E-mail: wangmaoyan@163.com

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	Mei-yan REN
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Cite this article:

REN Mei-yan,TANG Kuan-gang,XUE Min,GUO Hui-qin,WANG Mao-yan. Functional Analysis of the AmNAC3 Transcription Factor Gene from Ammopiptanthus mongolicus in Drought and Cold Resistances. China Biotechnology, 2019, 39(8): 32-39.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20190805 OR https://manu60.magtech.com.cn/biotech/Y2019/V39/I8/32

Fig.1 Water contents in the soil and seedlings and AmNAC3 expression levels under the drought and cold stresses (a) Relative water contents in soil and seedlings during the drought treatment period (b), (c) The expression levels of AmNAC3 under the drought and cold stress treatments

Fig.2 Identification patterns of the coding region, expression vector construction and RT-PCR detection of transgenic lines of AmNAC3

Fig.3 Water loss of detached leaves of the AmNAC3 transgenic Arabidopsis (a) Phenotypes of the detached leaves during dehydration for 0 to 7h (b) Water loss rates of the detached leaves

Fig.4 Stomatal aperture and ABI expression levels of AmNAC3 transgenic Arabidopsis leaves (a) Stomata observation of the leaves (b) Measurement of stomatal aperture (c) ABI expression levels


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