Analyses of diverse low alkaloid tobacco germplasm identify naturally occurring nucleotide variability contributing to reduced leaf nicotine accumulation

Burner, Nathaniel; McCauley, Abigail; Pramod, Sreepriya; Frederick, Jesse; Steede, Tyler; Kernodle, Sheri P.; Lewis, Ramsey S.

doi:10.1007/s11032-021-01274-5

Cited by 4 publications

(12 citation statements)

References 26 publications

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“…The low‐alkaloid TI313 line carrying nic1‐3 was backcrossed three times to Petit Havana SR‐I to remove most of the TI313 background. We confirmed that a 5‐bp deletion at MYC2a in TI313, which was reported during the course of this study (Burner et al., 2022), was not present through the repeated crossings by checking the sequences of the plants obtained after the second backcross.…”

Section: Methodssupporting

confidence: 89%

“…A deletion of ERF199 has been detected in nine tobacco germplasms with low‐nicotine traits: TI95, TI313, TI554, and TI1041 analyzed in this study and TI508, TI516, TI527, TI1211, and TI785 reported additionally by Burner et al. (2022). The positions and sizes of these individual ERF199 deletions were mostly distinct between the examined lines (Figure 4a, Figure S4), suggesting the occurrence of multiple alleles that possibly arose independently, indicating that ERF199 may be a mutational hotspot for low‐nicotine traits.…”

Section: Discussionsupporting

confidence: 63%

“…We designated the allele defined by this deletion in TI313 as nic1-3 (Figure 4a). To prepare a homozygous nic1-3 mutant with a clear background, we backcrossed T313 three times to tobacco Petit Havana SR-I and confirmed that the resulting lines after the second backcross do not harbor a 5-bp deletion in MYC2a recently reported to be present in TI313 (Burner et al, 2022).…”

Section: Identification Of Novel Low-nicotine Tobacco Lines and Their...mentioning

confidence: 55%

“…To expand the genetic resources of low‐nicotine phenotypes, we obtained seeds for 37 lines and confirmed that 11 of them were low‐nicotine lines based on the nicotine levels (<0.2 mg/g dry weight [DW] in Experiment 1, <1 mg/g DW in Experiment 2) in the leaves of in‐house‐grown plants (Figure S2). All the lines selected here were also evaluated as being low‐alkaloid accessions in an independent study (Burner et al., 2022).…”

Section: Resultsmentioning

confidence: 99%

“…In summary, we revealed an allelic series at the NIC1 and NIC2 loci across a number of tobacco germplasms and highlighted the significance of ERF189 and ERF199 in the regulation of the nicotine pathway (Hayashi et al., 2020). Although not the focus of this study, it is clear that other as yet undefined factors also contribute to low‐nicotine phenotypes (Burner et al., 2022), as alkaloid accumulation is under complex genetic control. It would be valuable to study further the low‐nicotine tobacco resources with the knowledge gained from the defined nic1 and nic2 mutations from this study.…”

Section: Discussionmentioning

confidence: 99%

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Natural and induced variations in transcriptional regulator genes result in low‐nicotine phenotypes in tobacco

et al. 2022

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In tobacco, the homologous ETHYLENE RESPONSE FACTOR (ERF) transcription factors ERF199 and ERF189 coordinate the transcription of multiple metabolic genes involved in nicotine biosynthesis. Natural alleles at the NIC1 and NIC2 loci greatly affect alkaloid accumulation and overlap with ERF199 and ERF189 in the tobacco genome, respectively. In this study, we identified several low-nicotine tobacco varieties lacking ERF199 or ERF189 from a tobacco germplasm collection. We characterized the sequence of these new nic1 and nic2 alleles, as well as the previously defined alleles nic1-1 and nic2-1. Moreover, we examined the influence of different nic alleles on alkaloid contents and expression levels of genes related to nicotine biosynthesis. We also demonstrated that the deletion of a distal genomic region attenuates ERF199 expression, resulting in a moderately negative effect on the alkaloid phenotype. Our study provides new insights into the regulation of nicotine biosynthesis and novel genetic resources to breed low-nicotine tobacco.

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Section: Methodssupporting

confidence: 89%

Section: Discussionsupporting

confidence: 63%

Section: Identification Of Novel Low-nicotine Tobacco Lines and Their...mentioning

confidence: 55%

Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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Natural and induced variations in transcriptional regulator genes result in low‐nicotine phenotypes in tobacco

et al. 2022

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Genetic regulation and manipulation of nicotine biosynthesis in tobacco: strategies to eliminate addictive alkaloids

Shoji,

Hashimoto,

Saito

2023

Journal of Experimental Botany

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Tobacco (Nicotiana tabacum L.) is a widely cultivated crop of the genus Nicotiana. Due to the highly addictive nature of tobacco products, tobacco smoking remains the leading cause of preventable death and disease. There is therefore a critical need to develop tobacco varieties with reduced or non-addictive nicotine levels. Nicotine and related pyridine alkaloids biosynthesized in the roots of tobacco plants are transported to the leaves, where they are stored in vacuoles as a defense against predators. Jasmonate, a defense-related plant hormone, plays a crucial signaling role in activating transcriptional regulators that coordinate the expression of downstream metabolic and transport genes involved in nicotine production. In recent years, substantial progress has been made in molecular and genomics research, revealing many metabolic and regulatory genes involved in nicotine biosynthesis. These advances have enabled us to develop tobacco plants with low or ultra-low nicotine levels through various methodologies, such as mutational breeding, genetic engineering, and genome editing. We review the recent progress on genetic manipulation of nicotine production in tobacco, which serves as an excellent example of plant metabolic engineering with profound social implications.

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Spectrophotometric and Chromatographic Determination of Alkaloids and Nicotine Contents in Lebanese Tobacco Leaves

Elchamieh,

Jaber,

Ibrahim

et al. 2024

Yüzüncü Yıl Üniversitesi Tarım Bilimleri Dergisi

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Nicotiana Tabacum; the annual herb; known as Tobacco from the Solanaceae family was known for its alkaloids and especially nicotine (NCT) content; smoking products, insecticides, anthelminthic activity and clinically proven therapeutics are examples of its uses. Herein two different methods were applied in an attempt to quantify the total alkaloids and NCT content in Nicotiana tabacum cultivated in Lebanon. Total alkaloids were investigated through the formation of a complex with bromocresol green under a simple spectrophotometric method. Whilst HPLC-DAD was the choice for the quantitation of NCT levels. The column was Lichrospher select B (5 µm, 250x4 mm), the temperature was set at 29 ⁰C and the wavelength at 260 nm. The mobile phase consisted of 2 M O-phosphoric acid and methanol (60:40, v/v) using isocratic elution at 1 mL/min. A linear relationship was proved under both instruments. The extraction yield of alkaloid totum ranges between 2.1 ± 0.25 and 6.8% ± 0.58 and alkaloids contents range from 12.14 ± 2.01 to 53.12 ± 4.54 mg of AE/g of extract for Ghandouriyeh and Danniye samples respectively. On the other side among the different areas which cultivated Tobacco in Lebanon, Danniye was found to have the highest NCT concentration of dry weight (2.64%) while Al-Hissa possesses the lowest content (0.75%). Even if the results were generally similar to other countries, the study showed a difference in values from one region to another.

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Analyses of diverse low alkaloid tobacco germplasm identify naturally occurring nucleotide variability contributing to reduced leaf nicotine accumulation

Cited by 4 publications

References 26 publications

Natural and induced variations in transcriptional regulator genes result in low‐nicotine phenotypes in tobacco

Natural and induced variations in transcriptional regulator genes result in low‐nicotine phenotypes in tobacco

Genetic regulation and manipulation of nicotine biosynthesis in tobacco: strategies to eliminate addictive alkaloids

Spectrophotometric and Chromatographic Determination of Alkaloids and Nicotine Contents in Lebanese Tobacco Leaves

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