AN EFFECTIVE METHOD OF RNA ISOLATION FROM<i>Fallopia multiflora</i>TUBEROUS ROOTS

Chen, Lei; Sheng, Shujing; Tan, Xue-Mei; Shen, Yanjun; Li, Hongqing; Zhao, S. J.

doi:10.1080/10826068.2011.566297

Cited by 5 publications

(6 citation statements)

References 15 publications

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“…Total RNA samples were isolated as described by Chen et al . (). qRT‐PCR analysis of CsGSTF1 was performed with SYBR Green dye (Sigma‐Aldrich) using an ABI 7500 Real‐Time PCR System with GAPDH (accession no.…”

Section: Methodsmentioning

confidence: 97%

A coupled role for CsMYB75 and CsGSTF1 in anthocyanin hyperaccumulation in purple tea

et al. 2018

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Cultivars of purple tea (Camellia sinensis) that accumulate anthocyanins in place of catechins are currently attracting global interest in their use as functional health beverages. RNA-seq of normal (LJ43) and purple Zijuan (ZJ) cultivars identified the transcription factor CsMYB75 and phi (F) class glutathione transferase CsGSTF1 as being associated with anthocyanin hyperaccumulation. Both genes mapped as a quantitative trait locus (QTL) to the purple bud leaf color (BLC) trait in F 1 populations, with CsMYB75 promoting the expression of CsGSTF1 in transgenic tobacco (Nicotiana tabacum). Although CsMYB75 elevates the biosynthesis of both catechins and anthocyanins, only anthocyanins accumulate in purple tea, indicating selective downstream regulation. As glutathione transferases in other plants are known to act as transporters (ligandins) of flavonoids, directing them for vacuolar deposition, the role of CsGSTF1 in selective anthocyanin accumulation was investigated. In tea, anthocyanins accumulate in multiple vesicles, with the expression of CsGSTF1 correlated with BLC, but not with catechin content, in diverse germplasm. Complementation of the Arabidopsis tt19-8 mutant, which is unable to express the orthologous ligandin AtGSTF12, restored anthocyanin accumulation, but did not rescue the transparent testa phenotype, confirming that CsGSTF1 did not function in catechin accumulation. Consistent with a ligandin function, transient expression of CsGSTF1 in Nicotiana occurred in the nucleus, cytoplasm and membrane. Furthermore, RNA-Seq of the complemented mutants exposed to 2% sucrose as a stress treatment showed unexpected roles for anthocyanin accumulation in affecting the expression of genes involved in redox responses, phosphate homeostasis and the biogenesis of photosynthetic components, as compared with non-complemented plants.

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Section: Methodsmentioning

confidence: 97%

A coupled role for CsMYB75 and CsGSTF1 in anthocyanin hyperaccumulation in purple tea

et al. 2018

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“…sinensis , CsF3′5′H1 (comp149726_c0) and three key F3′H genes ( CsF3′H1 , comp143838_c0; CsF3′H2 , comp147206_c0; CsF3′H3 , comp141172_c0) were selected for real time RT-PCR analysis, using RNA isolated from tea samples from thirteen varieties. RNA isolation was done according to a modified CTAB method described by Chen et al (2012) [ 27 ]. Primers were designed using Primer 3 software to amplify 200-241bp fragments for F3′Hs and 169 bp for F3′5′H1 selected from the RNA-Seq libraries ( Table 1 ).…”

Section: Methodsmentioning

confidence: 99%

Transcriptome Analysis Reveals Key Flavonoid 3′-Hydroxylase and Flavonoid 3′,5′-Hydroxylase Genes in Affecting the Ratio of Dihydroxylated to Trihydroxylated Catechins in Camellia sinensis

et al. 2015

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The ratio of dihydroxylated to trihydroxylated catechins (RDTC) is an important indicator of tea quality and biochemical marker for the study of genetic diversity. It is reported to be under genetic control but the underlying mechanism is not well understood. Flavonoid 3′-hydroxylase (F3′H) and flavonoid 3′,5′-hydroxylase (F3′5′H) are key enzymes involved in the formation of dihydroxylated and trihydroxylated catechins. The transcriptome and HPLC analysis of tea samples from Longjing43 and Zhonghuang2 under control and shading treatment were performed to assess the F3′H and F3′5′H genes that might affect RDTC. A total of 74.7 million reads of mRNA seq (2×101bp) data were generated. After de novo assembly, 109,909 unigenes were obtained, and 39,982 of them were annotated using 7 public databases. Four key F3′H and F3′5′H genes (including CsF3′5′H1, CsF3′H1, CsF3′H2 and CsF3′H3) were identified to be closely correlated with RDTC. Shading treatment had little effect on RDTC, which was attributed to the stable expression of these key F3′H and F3′5′H genes. The correlation of the coexpression of four key genes and RDTC was further confirmed among 13 tea varieties by real time PCR and HPLC analysis. The coexpression of three F3′H genes and a F3′5′H gene may play a key role in affecting RDTC in Camellia sinensis. The current results may establish valuable foundation for further research about the mechanism controlling catechin composition in tea.

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“…RNA isolation was done according to a modified CTAB method described by Chen et al (2012) [20]. Primers were designed using Primer 3 software to amplify 150–250 bp fragments for 36 transcripts selected from the RNA-Seq libraries as potential candidate genes (Table S1).…”

Section: Methodsmentioning

confidence: 99%

Transcriptome Analysis of Indole-3-Butyric Acid-Induced Adventitious Root Formation in Nodal Cuttings of Camellia sinensis (L.)

Wei

Wang

et al. 2014

PLoS ONE

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Tea (Camellia sinensis L.) is a popular world beverage, and propagation of tea plants chiefly depends on the formation of adventitious roots in cuttings. To better understand potential mechanisms involved in adventitious root formation, we performed transcriptome analysis of single nodal cuttings of C. sinensis treated with or without indole-3-butyric acid (IBA) using the Illumina sequencing method. Totally 42.5 million RNA-Seq reads were obtained and these were assembled into 59,931 unigenes, with an average length of 732 bp and an N50 of 1292 bp. In addition, 1091 differentially expressed unigenes were identified in the tea cuttings treated with IBA compared to controls, including 656 up- and 435 down-regulated genes. Further real time RT-PCR analysis confirmed RNA-Seq data. Functional annotation analysis showed that many genes were involved in plant hormone signal transduction, secondary metabolism, cell wall organization and glutathione metabolism, indicating potential contributions to adventitious rooting. Our study presents a global view of transcriptome profiles of tea cuttings in response to IBA treatment and provides new insights into the fundamental mechanisms associated with auxin-induced adventitious rooting. Our data will be a valuable resource for genomic research about adventitious root formation in tea cuttings, which can be used to improve rooting for difficult-to-root varieties.

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AN EFFECTIVE METHOD OF RNA ISOLATION FROMFallopia multifloraTUBEROUS ROOTS

Cited by 5 publications

References 15 publications

A coupled role for CsMYB75 and CsGSTF1 in anthocyanin hyperaccumulation in purple tea

A coupled role for CsMYB75 and CsGSTF1 in anthocyanin hyperaccumulation in purple tea

Transcriptome Analysis Reveals Key Flavonoid 3′-Hydroxylase and Flavonoid 3′,5′-Hydroxylase Genes in Affecting the Ratio of Dihydroxylated to Trihydroxylated Catechins in Camellia sinensis

Transcriptome Analysis of Indole-3-Butyric Acid-Induced Adventitious Root Formation in Nodal Cuttings of Camellia sinensis (L.)

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