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您的检索式：作者名="Thomas G.Ranney"
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| 1 | An optimized protocol for stepwise optimization of real-time RT-PCR analysis显示文摘Computational tool-assisted primer design for real-time reverse transcription(RT)PCR(qPCR)analysis largely ignores the sequence similarities between sequences of homologous genes in a plant genome.It can lead to false confidence in the quality of the designed primers,which sometimes results in skipping the optimization steps for qPCR.However,the optimization of qPCR parameters plays an essential role in the efficiency,specificity,and sensitivity of each gene’s primers.Here,we proposed an optimized approach to sequentially optimizing primer sequences,annealing temperatures,primer concentrations,and cDNA concentration range for each reference(and target)gene.Our approach started with a sequence-specific primer design that should be based on the single-nucleotide polymorphisms(SNPs)present in all the homologous sequences for each of the reference(and target)genes under study.By combining the efficiency calibrated and standard curve methods with the 2−ΔΔCt method,the standard cDNA concentration curve with a logarithmic scale was obtained for each primer pair for each gene.As a result,an R 2≥0.9999 and the efficiency(E)=100±5% should be achieved for the best primer pair of each gene,which serve as the prerequisite for using the 2^(−ΔΔCt) method for data analysis.We applied our newly developed approach to identify the best reference genes in different tissues and at various inflorescence developmental stages of Tripidium ravennae,an ornamental and biomass grass,and validated their utility under varying abiotic stress conditions.We also applied this approach to test the expression stability of six reference genes in soybean under biotic stress treatment with Xanthomonas axonopodis pv.glycines(Xag).Thus,these case studies demonstrated the effectiveness of our optimized protocol for qPCR analysis. | Fangzhou Zhao Nathan A.Maren Pawel Z.Kosentka Ying-Yu Liao Hongyan Lu James R.Duduit Debao Huang Hamid Ashrafi Tuanjie Zhao Alejandra I.Huerta Thomas G.Ranney Wusheng Liu | 2021 | Horticulture Research2021，8，1： | 2 |
| 2 | New opportunities for using WUS/BBM and GRF-GIF genes to enhance genetic transformation of ornamental plants显示文摘Broad application of plant transformation remains challenging because the efficiency of plant regeneration and regeneration-based transformation in many plant species is extremely low.Many species and genotypes are not responsive to traditional hormone-based regeneration systems.This regeneration recalcitrance hampers the application of many technologies such as micropropagation,transgenic breeding,and gene editing in various plant species,including ornamental flowers,shrubs,and trees.Various developmental genes have long been studied for their ability to improve plant meristematic induction and regeneration.Lately,it was demonstrated that the combined and refined expression of morphogenic regulator genes WUSCHEL and BABY BOOM could alleviate their pleiotropic effects and permit transformation in recalcitrant monocots.Moreover,ectopic expression of plant growth-regulating factors(GRFs)alone or in combination with GRF-interacting factors(GIFs)improved the regeneration and transformation of dicot and monocot species.Fine-tuning the expression of these genes provides new opportunities to improve transformation efficiencies and facilitate the application of new breeding technologies in ornamental plants. | Hui Duan Nathan A.Maren Thomas G.Ranney Wusheng Liu | 2022 | Ornamental Plant Research2022，2，1： | 0 |
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