Large-scale development of SSR markers in tobacco and construction of a linkage map in flue-cured tobacco

Tong, Zhijun; Xiao, Bingguang; Fangchan, Jiao; Fang, Dunhuang; Zeng, Jianmin; Xingfu, Wu; Chen, Xuejun; Yang, Jiankang; Li, Yongping

doi:10.1270/jsbbs.15129

Cited by 31 publications

(25 citation statements)

References 25 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In total, 13,645 simple sequence repeat (SSR) markers were screened for identifying the segregating markers in the NIC1-locus region. PCR of the SSR markers, PCR product separation and visualization were performed as described previously (Tong et al, 2016). Genotyping of the F 2 population were conducted using the linkage markers and the Kosambi mapping function was used to transform the recombination frequency to genetic distance (cM).…”

Section: Field Trail and Genetic Mappingmentioning

confidence: 99%

Unravel the Mystery ofNIC1-locus on Nicotine Biosynthesis Regulation in Tobacco

Sui

Xie

Tong

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

ABSTRACTBackgroundNicotine biosynthesis is mainly regulated by jasmonate (JA) signaling cascade in Nicotiana tabacum. As an allotetraploid species, the regulation of nicotine biosynthesis has been genetically verified via two unlinked NIC loci (named as NIC1 and NIC2) which are possibly originated from its two ancestral diploids. Previously, a N. tomentosiformis originated ethylene response factor (ERF) gene cluster was identified as the NIC2-locus which has been demonstrated positively regulates nicotine accumulation in N. tabacum.ResultsHere, we describe the genetic mapping of NIC1-locus, the major nicotine regulatory locus, by using a NIC1-locus segregating population through bulked segregant analysis. We identified two linkage marker TM23004 and TM22038 were delimited the NIC1-locus within a ~34.3-Mb genomic region at pseudochromosome 07 of tobacco genome. Genomic scan within this region revealed a NIC2-like locus ERF gene cluster exist in. To verify this ERF gene cluster is the genetically called “NIC1-locus”, different functional experiments based on most of the ERFs in regulating nicotine biosynthesis and their influences on alkaloid accumulations have been carried out. Collinearity analysis showed that NIC1-locus ERF genes are originated from N. sylvestris and exclusively expressed in root tissues. In addition, transcriptomic results indicate that NIC1-locus ERF genes are coexpressed with the NIC2-locus ERF genes and other nicotine biosynthetic genes and regulators after JA induction. Furthermore, the suppressed expression of four ERFs of the NIC1-locus genes corresponding with decreased NtPMT and NtQPT expression in NtMYC2-RNAi lines indicates the selected NIC1-locus ERFs function in downstream of NtMYC2 in the JA signaling cascades. In the meanwhile, the alkaloid levels are also determined by the amplitude of the four ERF gene expressions in both wild type and LA mutant. Additionally, in vitro binding assays, transient activation assays, and ectopic expression in transgenic plants demonstrate that these ERF genes are able to bind the GCC-box elements residing in the step-limiting gene promoters (such as NtPMT2, NtQPT2) and functional redundant but quantitatively transactivate nicotine biosynthetic gene expression. For nic1-locus mutation, two different sizes of deletions (nic1-S and nic1-B) were identified which occurred at the surrounding regions of the NIC1-locus gene cluster, which might disrupt, to some extent, chromosomal microenvironment and change gene expression around the deletion regions (including NIC1-locus ERFs), resulting in the decreased expression levels of NIC1-locus ERFs (such as NtERF199) and reduced alkaloid accumulation in the nic1-locus mutant.ConclusionsOur findings not only provide insight in to the mechanism of the NIC1-locus ERFs in the regulatory network of nicotine biosynthesis, but also unraveled the theoretical basis of the nic1-locus mutation in low nicotine mutant. These functional verified NIC1-locus ERF genes can be further used as potential target(s) for ethyl methanesulfonate-based mutagenesis to manipulate nicotine level in tobacco variety in tobacco breeding program.

show abstract

Section: Field Trail and Genetic Mappingmentioning

confidence: 99%

Unravel the Mystery ofNIC1-locus on Nicotine Biosynthesis Regulation in Tobacco

Sui

Xie

Tong

et al. 2020

Preprint

Self Cite

View full text Add to dashboard Cite

show abstract

“…Flue-cured tobacco is the most important type of tobacco and accounts for 66% of all types of tobacco produced globally [28]. The evaluation of FCTL quality is largely based on sensory evaluations of quality and style.…”

Section: Introductionmentioning

confidence: 99%

A predictive model for the sensory aroma characteristics of flue-cured tobacco based on a back-propagation neural network

Chen

Kawaguchi

et al. 2020

SN Appl. Sci.

View full text Add to dashboard Cite

In this study, aroma quality, aroma quantity, diffusiveness and offensive odor predictions were modeled using a back propagation neural network (BPNN). The model was built from chemical components and sensory quality data features of flue-cured tobacco leaf samples. The results showed that the BPNN model adapted the optimized hidden layer well and had excellent generalization ability and robustness. The coefficients of determination (R 2) for the indices of tobacco aroma characteristics based on the BPNN model, namely, aroma quality, aroma quantity, diffusiveness and offensive odor, were all above 0.7. The accuracies of the model parameters (mean squared error, regression mean standard error , and mean absolute error) were better than those of the model built using stepwise regression analysis. These results indicate that the BPNN prediction model is reliable and can accurately predict the sensory aroma characteristic qualities of flue-cured tobacco leaf.

show abstract

“…Therefore, molecular marker-based methods represent an emerging trend in rapid DUS testing [2], [7], [11], [12], [13]. Of the numerous molecular marker technologies available, simple sequence repeats (SSR) analysis is considered ideal for the DUS testing of new varieties [8], [14], [15], [16], [17], [18] and the ngerprinting of standard crop varieties [10], [11], [12] due to multiple associated advantages, such as the abundance, high polymorphism, and co-dominance of SSR markers [19], [20] and the stability, repeatability, and simple operational procedures involved in SSR analysis [10], [21], [22].…”

Section: Introductionmentioning

confidence: 99%

Genetic diversity and fingerprinting of 33 standard flue-cured tobacco varieties for use in distinctness, uniformity, and stability testing

Geng

Cheng

et al. 2020

Preprint

View full text Add to dashboard Cite

Background: At present, the distinctness, uniformity, and stability (DUS) testing of flue-cured tobacco ( Nicotiana tabacum L.) depends on field morphological identification, which is problematic in that it is intensive, time-consuming, and susceptible to environmental impacts. In order to improve the efficiency and accuracy of tobacco DUS testing, the development of a molecular marker-based method for genetic diversity identification is urgently needed. Results: In total, 91 simple sequence repeats (SSR) markers with clear and polymorphic amplification bands were obtained with polymorphism information content, Nei index, and Shannon information index values of 0.3603, 0.4040, and 0.7228, respectively. Clustering analysis showed that the 33 study varieties, which are standard varieties for flue-cured tobacco DUS testing, could all be distinguished from one another. Further analysis showed that a minimum of 25 markers were required to identify the genetic diversity of these varieties. Following the principle of two markers per linkage group, 48 pairs of SSR markers were selected. Correlation analysis showed that the genetic relationships revealed by the 48 SSR markers were consistent with those found using the 91 SSR markers. Conclusions: The genetic fingerprints of the 33 standard varieties of flue-cured tobacco were constructed using 48 SSR markers, and an SSR marker-based identification technique for new tobacco varieties was developed. This study provides a reliable technological approach for determining the novelty of new tobacco varieties and offers a solid technical basis for the accreditation and protection of new tobacco varieties.

show abstract

Large-scale development of SSR markers in tobacco and construction of a linkage map in flue-cured tobacco

Cited by 31 publications

References 25 publications

Unravel the Mystery ofNIC1-locus on Nicotine Biosynthesis Regulation in Tobacco

Unravel the Mystery ofNIC1-locus on Nicotine Biosynthesis Regulation in Tobacco

A predictive model for the sensory aroma characteristics of flue-cured tobacco based on a back-propagation neural network

Genetic diversity and fingerprinting of 33 standard flue-cured tobacco varieties for use in distinctness, uniformity, and stability testing

Contact Info

Product

Resources

About