WRINKLED1 homologs highly and functionally express in oil-rich endosperms of oat and castor

Zheng, Yanan; Liu, Xiangling; Li, Na; Du, Can; Wang, Kai; Wang, Zhonghua; Hu, Yin-Gang; Zhang, Meng

doi:10.1016/j.plantsci.2019.110193

Cited by 29 publications

(26 citation statements)

References 48 publications

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“…Their oil content and fatty acid composition were determined by the modified SAST method. Results showed that oil content of T 1 seeds increased from 5 μg/seed to nearly 8 μg/seed, which is consistent with the improvement in the transgenic T 3 seeds in the previous study (Figure 5a in this study and Figure 5a of Yang et al, 2019). These results indicate that quantifying oil content of T 1 seeds by the five-seed modified SAST method is valid to predict the variation of oil contents in transgene experiments.…”

Section: Application Of the Sast Or Modified Sast To Phenotype Fattsupporting

confidence: 91%

“…However, it is worth noting that the composition of transmethylation solution and parameters of incubation varies among previous studies. For examples, concentration of catalyst H 2 SO 4 in transmethylation solution range from 2.5% to 5% (Epp and Pollard, 1993, Lee et al, 2017, Yang et al, 2019. Toluene is a cosolvent of triacylglycerol and methanol and 30% toluene is added in transmethylation solution or it may be not included (Zhang et al, 2009, Lee et al, 2017.…”

Section: Discussionmentioning

confidence: 99%

“…RcWRI1 was cloned from castor been and seed-specific overexpression of RcWRI1 in Arabidopsis increased oil contents of T 3 seeds (Yang et al, 2019). In this study, the same construct was transferred into Arabidopsis and T 1 seeds were identified by their DsRed fluorescence.…”

Section: Application Of the Sast Or Modified Sast To Phenotype Fattmentioning

confidence: 98%

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Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds

Ohlrogge

et al. 2020

Plant Direct

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Arabidopsis is wildly used as a model plant and camelina is increasingly used for oilseed research and applications. Although the Arabidopsis genome has been sequenced for two decades, the functions of many lipid‐related genes and their regulators have not been well characterized. Improvements in the efficiency and accuracy of gene investigations are key to effective discovery of gene function and downstream bioengineering of plant oil quantity and quality. In this study, a visible marker was used to quickly identify transgenic T1 seeds and a method has been developed to phenotype fatty acid compositions and oil content of single T1 seeds. A whole seed direct transmethylation method was first optimized with multiple seeds and incubation at 85°C for 2 hours in a transmethylation solvent (5% H2SO4 in methanol with 30% toluene cosolvent) is recommended. Based on this method, a single Arabidopsis seed mini‐transmethylation (SAST) method has been established in a 1.5 ml GC sample vial with 200 μl transmethylation solvent. Characteristics of the method were evaluated and it was used to phenotype transgenic T1 seeds expressing AtFAD2 or RcWRI1. Our results indicate that fatty acid composition of T1 individual seeds are consistent with those of pools of multiple seeds from higher generations. However, oil content per individual seed varied substantially and therefore pooling five seeds is recommended for phenotyping oil content of T1 seeds. Additionally, a whole camelina single‐seed direct transmethylation was evaluated and results confirm its feasibility. The suitability of partial seed analysis of camelina was investigated but variation in composition of different seed tissues limits this approach.

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Section: Application Of the Sast Or Modified Sast To Phenotype Fattsupporting

confidence: 91%

Section: Discussionmentioning

confidence: 99%

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Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds

Ohlrogge

et al. 2020

Plant Direct

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“…The fatty acid was measured following Gas Chromatography (GC) measurement as described previously (Li et al, 2006;Yang et al, 2019). About 10 mg dry tissue powder was esterified in 2 mL methanol with 2.5% (v/v) H 2 SO 4 , and 50 m g triheptadecanoin was used as internal standard.…”

Section: Fatty Acid Measurementmentioning

confidence: 99%

Single Nucleotide Mutagenesis of the TaCHLI Gene Suppressed Chlorophyll and Fatty Acid Biosynthesis in Common Wheat Seedlings

Wang

Zhang

et al. 2020

Front. Plant Sci.

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Wheat (Triticum aestivum L.) is one of the most important crops in the world. Chlorophyll plays a vital role in plant development and crop improvement and further determines the crop productivity to a certain extent. The biosynthesis of chlorophyll remains a complex metabolic process, and fundamental biochemical discoveries have resulted from studies of plant mutants with altered leaf color. In this study, we identified a chlorophyll-deficiency mutant, referred to as chli, from the wheat cultivar Shaannong33 that exhibited an obvious pale-green leaf phenotype at the seedling stage, with significantly decreased accumulation of chlorophyll and its precursors, protoporphyrin IX and Mgprotoporphyrin IX. Interestingly, a higher protoporphyrin IX to Mg-protoporphyrin IX ratio was observed in chli. Lipid biosynthesis in chli leaves and seeds was also affected, with the mutant displaying significantly reduced total lipid content relative to Shaanong33. Genetic analysis indicated that the pale-green leaf phenotype was controlled by a single pair of recessive nuclear genes. Furthermore, sequence alignment revealed a single-nucleotide mutation (G664A) in the gene TraesCS7A01G480700.1, which encodes subunit I of the Mg-chelatase in plants. This single-nucleotide mutation resulted in an amino acid substitution (D221N) in the highly conserved domain of subunit I. As a result, mutant protein Tachli-7A lost the ability to interact with the normal protein TaCHLI-7A, as assessed by yeast two-hybrid assay. Meanwhile, Tachli-7A could not recover the chlorophyll deficiency phenotype of the Arabidopsis thaliana SALK_050029 mutant. Furthermore, we found that in Shaannong33, the protoporphyrin IX to Mgprotoporphyrin IX ratio was growth state-dependent and insensitive to environmental change. Overall, the mutation in Tachli-7A impaired the function of Mg-chelatase and blocked the conversion of protoporphyrin IX to Mg-protoporphyrin IX. Based on our results, the chli mutant represents a potentially useful resource for better understanding chlorophyll and lipid biosynthetic pathways in common wheat.

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“…Meanwhile, as the largest tissue of oat seeds, endosperms have a unique characterization among monocots that most lipids of oat seeds are stored in its endosperms. This characterization arouses researchers' increasing interests in the oil accumulation mechanism of oat endosperms [13,14]. Thus, the molecular biological analyses of oat seeds and endosperms are also required, and seven stages of developing seeds and corresponding endosperms were collected as samples according to Ekman et al [7].…”

Section: Selection Of Oat Tissues and Expression Stabilities Of Candimentioning

confidence: 99%

Evaluation of duplicated reference genes for quantitative real-time PCR analysis in genome unknown hexaploid oat (Avena sativa L.)

Yang

Wang

Aziz

et al. 2020

Preprint

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Background: Oat (Avena sativa L.), a hexaploid crop with unknown genome, has valuable nutritional, medicinal and pharmaceutical uses. However, no suitable RGs (reference genes) for qPCR (quantitative real-time PCR) has been documented for oat yet. Single-copy gene is often selected as RG, which is challengeable or impactable in unexplored polyploids.Results: In this study, eleven candidate RGs, including four duplicated genes, were selected from oat transcriptome. The stability and the optimal combination of these candidate RGs were assessed in 18 oat samples by using four statistical algorithms including the ΔCt method, geNorm, NormFinder and BestKeeper. The most stable RGs for “all samples”, “shoots and roots of seedlings”, “developing seeds” and “developing endosperms” were EIF4A (Eukaryotic initiation factor 4A-3), UBC21 (Ubiquitin-Conjugating Enzyme 21), EP (Expressed protein) and EIF4A respectively. Among these RGs, UBC21 was a four-copy duplicated gene. The reliability was validated by the expression patterns of four various genes normalized to the most and the least stable RGs in different sample sets.Conclusions: Results provide a proof of concept that the duplicated RG is feasible for qPCR in polyploids. To our knowledge, this study is the first systematic research on the optimal RGs for accurate qPCR normalization of gene expression in different organs and tissues of oat.

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WRINKLED1 homologs highly and functionally express in oil-rich endosperms of oat and castor

Cited by 29 publications

References 48 publications

Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds

Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds

Single Nucleotide Mutagenesis of the TaCHLI Gene Suppressed Chlorophyll and Fatty Acid Biosynthesis in Common Wheat Seedlings

Evaluation of duplicated reference genes for quantitative real-time PCR analysis in genome unknown hexaploid oat (Avena sativa L.)

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WRINKLED1 homologs highly and functionally express in oil-rich endosperms of oat and castor

Cited by 29 publications

References 48 publications

Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T1 Arabidopsis and camelina seeds

Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T1 Arabidopsis and camelina seeds

Single Nucleotide Mutagenesis of the TaCHLI Gene Suppressed Chlorophyll and Fatty Acid Biosynthesis in Common Wheat Seedlings

Evaluation of duplicated reference genes for quantitative real-time PCR analysis in genome unknown hexaploid oat (Avena sativa L.)

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Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds

Accelerating gene function discovery by rapid phenotyping of fatty acid composition and oil content of single transgenic T₁ Arabidopsis and camelina seeds