Ectopic Expression of Pumpkin NAC Transcription Factor CmNAC1 Improves Multiple Abiotic Stress Tolerance in Arabidopsis

Cao, Haishun; Wang, Li; Nawaz, Muhammad Azher; Niu, Mengliang; Sun, Jingyu; Xie, Jin; Kong, Qiusheng; Huang, Yuan; Cheng, Fei; Bie, Zhilong

doi:10.3389/fpls.2017.02052

Cited by 41 publications

(37 citation statements)

References 62 publications

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“…The T1 generations were said to be the transgenic tobacco lines that contained a single insertion, whose seeds were grown in the same conditions as described earlier. After three weeks, the seedlings were transferred to a hydroponic system and two weeks later, the seedlings of the transgenic tobacco were exposed to 100 mM NaCl [ 34 ], 15% PEG-6000 ( m / v ) [ 35 ] and 100 mM H 2 O 2 [ 36 ] in half strength Hoagland’s nutrient solution (24 h) for simulating salt, drought and oxidative stress, respectively. For cold stress, the seedlings were cultivated in half strength Hoagland’s solution at 4 °C [ 37 ].…”

Section: Methodsmentioning

confidence: 99%

Cloning and Functional Analysis of Phosphoethanolamine Methyltransferase Promoter from Maize (Zea mays L.)

Niu

Gou

Han

et al. 2018

IJMS

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Betaine, a non-toxic osmoprotectant, is believed to accumulate considerably in plants under stress conditions to maintain the osmotic pressure and promote a variety of processes involved in growth and development. Phosphoethanolamine N-methyltransferase (PEAMT), a key enzyme for betaine synthesis, is reported to be regulated by its upstream promoter. In the present investigation, by using the transgenic approach, a 1048 bp long promoter region of ZmPEAMT gene from Zea mays was cloned and functionally characterized in tobacco. Computational analysis affirmed the existence of abiotic stress responsive cis-elements like ABRE, MYC, HST, LST etc., as well as pathogen, wound and phytohormone responsive motifs. For transformation in tobacco, four 5′-deletion constructs of 826 bp (P2), 642 bp (P3), 428 bp (P4) and 245 bp (P5) were constructed from the 1048 bp (P1) promoter fragment. The transgenic plants generated through a single event exhibited a promising expression of GUS reporter protein in the leaf tissues of treated with salt, drought, oxidative and cold stress as well as control plants. The GUS expression level progressively reduced from P1 to P5 in the leaf tissues, whereas a maximal expression was observed with the P3 construct in the leaves of control plants. The expression of GUS was noted to be higher in the leaves of osmotically- or salt-treated transgenic plants than that in the untreated (control) plants. An effective expression of GUS in the transgenic plants manifests that this promoter can be employed for both stress-inducible and constitutive expression of gene(s). Due to this characteristic, this potential promoter can be effectively used for genetic engineering of several crops.

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Section: Methodsmentioning

confidence: 99%

Cloning and Functional Analysis of Phosphoethanolamine Methyltransferase Promoter from Maize (Zea mays L.)

Niu

Gou

Han

et al. 2018

IJMS

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“…Examining water loss rate from detached leaves/rosettes has been widely used as important, reliable indicator to rapidly predict the plant water status and performance under drought conditions [53][54][55]. A great number previous studies included this tool in evaluating the performance of various transgenic plants, with the tracking of water loss rates over the course duration of 3 h [56,57], 4 h [58,59], 5 h and longer [52,60]. Upon being excised, water loss is indicated to be achieved via stomatal transpiration within the first hour followed by cuticular transpiration and residual transpiration (i.e., from incompletely closed stomata) in subsequent hours [61].…”

Section: Discussionmentioning

confidence: 99%

Heterologous Expression of a Soybean Gene RR34 Conferred Improved Drought Resistance of Transgenic Arabidopsis

Nghia

Chuong

Hoang

et al. 2020

Plants

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Two-component systems (TCSs) have been identified as participants in mediating plant response to water deficit. Nevertheless, insights of their contribution to plant drought responses and associated regulatory mechanisms remain limited. Herein, a soybean response regulator (RR) gene RR34, which is the potential drought-responsive downstream member of a TCS, was ectopically expressed in the model plant Arabidopsis for the analysis of its biological roles in drought stress response. Results from the survival test revealed outstanding recovery ratios of 52%–53% in the examined transgenic lines compared with 28% of the wild-type plants. Additionally, remarkedly lower water loss rates in detached leaves as well as enhanced antioxidant enzyme activities of catalase and superoxide dismutase were observed in the transgenic group. Further transcriptional analysis of a subset of drought-responsive genes demonstrated higher expression in GmRR34-transgenic plants upon exposure to drought, including abscisic acid (ABA)-related genes NCED3, OST1, ABI5, and RAB18. These ectopic expression lines also displayed hypersensitivity to ABA treatment at germination and post-germination stages. Collectively, these findings indicated the ABA-associated mode of action of GmRR34 in conferring better plant performance under the adverse drought conditions.

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“…Manuscript to be reviewed 2017; Obrero et al, 2011;Tashiro et al, 2016;Niu et al, 2017) and have been used for gene expression studies in cucumber (Wang et al, 2009;Migocka and Papierniak, 2011;Warzybock and Migocka, 2013). The genes UFP (ubiquitin), EF-1A (elongation factor), PRL36aA (60S ribosomal protein L36a/L44), PP2A (protein phosphatase) and CACS (clathrin adaptor complexes medium submit family protein) have been used for reliable normalization in different experimental sets in zucchini (Cucurbita pepo) (Obrero et al, 2011), and these reference genes have been successfully applied to both cucumber and pumpkin in specific environments, including powdery mildew, salinity, cold, dehydration, H 2 O 2 , and abscisic acid (ABA) treatments (Berg et al, 2015;Cao et al, 2017;Reda et al, 2018). Unfortunately, no single reference gene has been confirmed that exhibits uniform and stable expression under different experimental conditions.…”

Section: Introductionmentioning

confidence: 99%

Peer Review #3 of "Selection of reference genes for quantitative real-time PCR analysis in cucumber (Cucumis sativus L.), pumpkin (Cucurbita moschata Duch.) and cucumber–pumpkin grafted plants (v0.1)"

Fahrentrapp¹

2019

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Background: Quantitative real-time PCR (qRT-PCR) is a commonly used high-throughput technique to measure mRNA transcript levels. The accuracy of this evaluation of gene expression depends on the use of optimal reference genes. Cucumber-pumpkin grafted plants, made by grafting a cucumber scion onto pumpkin rootstock, are superior to either parent plant, as grafting conveys many advantages. However, although many reliable reference genes have been identified in both cucumber and pumpkin, none have been obtained for cucumber-pumpkin grafted plants. Methods: In this work, 12 candidate reference genes, including 8 traditional genes and 4 novel genes identified from our transcriptome data, were selected to assess their expression stability. Their expression levels in 25 samples, including 3 cucumber and 3 pumpkin samples from different organs, and 19 cucumber-pumpkin grafted samples from different organs, conditions, and varieties, were analyzed by qRT-PCR, and the stability of their expression was assessed by the comparative ΔCt method, geNorm, NormFinder, BestKeeper, and RefFinder. Results: The results showed that the most suitable reference gene varied dependent on the organs, conditions, and varieties. CACS and 40SRPS8 were the most stable reference genes for all samples in our research. TIP41 and CACS showed the most stable expression in different cucumber organs, TIP41 and PP2A were the optimal reference genes in pumpkin organs, and CACS and 40SRPS8 were the most stable genes in all grafted cucumber samples. However, the optimal reference gene varied under different conditions. CACS and 40SRPS8 were the best combination of genes in different organs of cucumber-pumpkin grafted plants, TUA and RPL36Aa were the most stable in the graft union under cold stress, LEA26 and ARF showed the most stable expression in the graft union during the healing process, and TIP41 and PP2A were the most stable across different varieties of cucumber-pumpkin grafted plants. The use of LEA26, ARF and LEA26+ARF as reference genes were further verified by analyzing the expression levels of csaCYCD3;1, csaRUL, cmoRUL, and cmoPIN in the graft union at different time points after grafting. Discussion: This work is the first report of appropriate reference genes in grafted cucumber plants and provides useful information for the study of gene expression and molecular mechanisms in cucumber-pumpkin grafted plants.

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Ectopic Expression of Pumpkin NAC Transcription Factor CmNAC1 Improves Multiple Abiotic Stress Tolerance in Arabidopsis

Cited by 41 publications

References 62 publications

Cloning and Functional Analysis of Phosphoethanolamine Methyltransferase Promoter from Maize (Zea mays L.)

Cloning and Functional Analysis of Phosphoethanolamine Methyltransferase Promoter from Maize (Zea mays L.)

Heterologous Expression of a Soybean Gene RR34 Conferred Improved Drought Resistance of Transgenic Arabidopsis

Peer Review #3 of "Selection of reference genes for quantitative real-time PCR analysis in cucumber (Cucumis sativus L.), pumpkin (Cucurbita moschata Duch.) and cucumber–pumpkin grafted plants (v0.1)"

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