Isolation and characterization of a high-affinity ammonium transporter ApAMT1;1 in alligatorweed

Guo, Xiaotong; Sheng, Yuting; Yang, Shaoyu; Han, Lei; Gao, Yachao; Zhang, Kai; Cheng, Jieshan; Zhang, Hongxia; Song, Zhizhong; Su, Yanhua

doi:10.1007/s10725-019-00537-8

Cited by 2 publications

(6 citation statements)

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“…Mutant. We further carried out functional determination of PpeAMT3;4 via complementation of the yeast mutant 31019b [6,13,17,30]. Results showed that yeast cells harboring pYES2 or pYES2-PpeAMT3;4 grew well, with similar growth status, on the YNB medium that contains 2 mM Arg as the sole N source (Figure 5).…”

Section: Functional Determination Of Ppeamt1;1 In Yeastmentioning

confidence: 99%

“…The recombinant plasmid pYES2-AMT3;4 was constructed by cloning the CDS region of the PpeAMT3;4 gene into the pYES2 expression vector [17], using the primer pairs (Table 1, Kpn I site was introduced and underlined in the forward primer, Not I site was introduced and underlined in the reverse primer). The yeast strain 31019b (MATa mepl△ mep2△::LEU2 mep3△::KanMX2 ura3, [7,13,17]) was transformed with pYES2 harboring the CDS of PpeAMT3;4. Yeast complementation tests were carried out as described in previous reports [7,13,17].…”

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

“…The yeast strain 31019b (MATa mepl△ mep2△::LEU2 mep3△::KanMX2 ura3, [7,13,17]) was transformed with pYES2 harboring the CDS of PpeAMT3;4. Yeast complementation tests were carried out as described in previous reports [7,13,17]. Yeast strain 31019b was transformed with pYES2 or pYES2-AMT3;4, respectively.…”

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

“…15 N Uptake Kinetics Assay. 15 N labeled NH 4 + uptake assays were carried out according to the description of Guo et al [17]. Yeast cells transformed with pYES2-MT3;4 or pYES2 were grown in the YNB liquid medium, supplemented with 15…”

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

“…The first plant AMT family gene was observed in Arabidopsis [7], and AtAMT1;1, AtAMT1;2, and AtAMT1;3 successfully restored the normal growth of the yeast mutant that is deficient in NH 4 + uptake systems. Subsequent studies of AMT transporters have been identified in diverse plant species, including Lotus japonicus (L. japonicus) [8,9], Lycopersicon esculentum (L. esculentum) [10][11][12], Populus trichocarpa (P. trichocarpa) [13], Oryza sativa (O. sativa) [14,15], Triticum aestivum (T. aestivum) [16], Alternanthera philoxeroides (A. philoxeroides) [17], and so on, which have been characterized in Xenopus oocytes or yeast mutant. The AMT family transporters can be divided into two subfamilies: AMT1 and AMT2 [3][4][5].…”

Section: Introductionmentioning

confidence: 99%

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Cloning and Functional Determination of Ammonium Transporter PpeAMT3;4 in Peach

You

Wang

et al. 2020

BioMed Research International

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Ammonium (NH4+) plays key roles in plant growth, development, fruit quality, and yield. In plants, NH4+ uptake and transport are facilitated by NH4+ transporters (AMT). However, molecular mechanisms and physiological functions of type-II AMT (AMT2) transporters in fruit trees are still unclear, especially in peach. In this study, we cloned and characterized an AMT2 family gene from peach, PpeAMT3;4, and determined its function in yeast mutant. Expression analysis showed that PpeAMT3;4 was majorly expressed in peach roots and significantly decreased by NH4+ excess but had no response to NH4+ deficiency. Functional determination and 15nitrogen-labeled NH4+ uptake assay in yeast cells implied that PpeAMT3;4 was a typical high-affinity transporter, with a K m value of 86.3 μM, that can uptake external NH4+ in yeast cells. This study provides gene resources to uncover the biological function of AMT2 transporters and reveals molecular basis for NH4+ uptake and nitrogen (N) nutrition mechanisms in fruit trees.

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Section: Functional Determination Of Ppeamt1;1 In Yeastmentioning

confidence: 99%

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

Section: Qrt-pcr Assays Specific Expression Primers Formentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Cloning and Functional Determination of Ammonium Transporter PpeAMT3;4 in Peach

You

Wang

et al. 2020

BioMed Research International

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Genome-Wide Identification and Characterization of Ammonium Transporter (AMT) Genes in Rapeseed (Brassica napus L.)

Dai

Han²,

Walk

et al. 2023

Genes

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Ammonium transporters (AMTs) are plasma membrane proteins mediating ammonium uptake and transport. As such, AMTs play vital roles in ammonium acquisition and mobilization, plant growth and development, and stress and pathogen defense responses. Identification of favorable AMT genotypes is a prime target for crop improvement. However, to date, systematic identification and expression analysis of AMT gene family members has not yet been reported for rapeseed (Brassica napus L.). In this study, 20 AMT genes were identified in a comprehensive search of the B. napus genome, 14 members of AMT1 and 6 members of AMT2. Tissue expression analyses revealed that the 14 AMT genes were primarily expressed in vegetative organs, suggesting that different BnaAMT genes might function in specific tissues at the different development stages. Meanwhile, qRT-PCR analysis found that several BnaAMTs strongly respond to the exogenous N conditions, implying the functional roles of AMT genes in ammonium absorption in rapeseed. Moreover, the rapeseed AMT genes were found to be differentially regulated by N, P, and K deficiency, indicating that crosstalk might exist in response to different stresses. Additionally, the subcellular localization of several BnaAMT proteins was confirmed in Arabidopsis protoplasts, and their functions were studied in detail by heterologous expression in yeast. In summary, our studies revealed the potential roles of BnaAMT genes in N acquisition or transportation and abiotic stress response and could provide valuable resources for revealing the functionality of AMTs in rapeseed.

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Isolation and characterization of a high-affinity ammonium transporter ApAMT1;1 in alligatorweed

Cited by 2 publications

References 53 publications

Cloning and Functional Determination of Ammonium Transporter PpeAMT3;4 in Peach

Cloning and Functional Determination of Ammonium Transporter PpeAMT3;4 in Peach

Genome-Wide Identification and Characterization of Ammonium Transporter (AMT) Genes in Rapeseed (Brassica napus L.)

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