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        Genome‑wide identification of GMP genes in Rosaceae and functional characterization of FaGMP4 in strawberry (Fragaria × ananassa)

        Yuanxiu Lin,Jiahao Zhang,Lintai Wu,Yunting Zhang,Qing Chen,Mengyao Li,Yong Zhang,Ya Luo,Yan Wang,Xiaorong Wang,Haoru Tang 한국유전학회 2021 Genes & Genomics Vol.43 No.6

        Background GDP-D-mannose pyrophosphorylase (GMP) is one of the key enzymes determining ascorbic acid (AsA) biosynthesis. However, little information about GMP genes is currently available for the Rosaceae species, especially in the AsA-riched cultivated octoploid strawberry (Fragaria × ananassa). Objective To identify the all the GMP genes in Rosaceae, as well as the predominant homologues and the role of GMP genes in strawberry AsA accumulation. Methods In the present study, we performed genome-wide identifcation and comprehensive analysis of the duplicated GMP genes in strawberry and other Rosaceae species by bioinformatics methods, the expression of the GMP genes from cultivated strawberry (Fragaria × ananassa, FaGMP) was specifcally analyzed by qPCR. Finally, the FaGMP4 was transiently overexpressed in strawberry to estimate the role of GMP in regulating AsA accumulation in strawberry. Results As results, a total of 28 GMP genes were identifed in the fve Rosaceae species. The origins of duplication events analysis suggested that most GMP duplications in Rosaceae species were generated from whole genome duplication (WGD). The Ka/Ks ratio suggested that FaGMP genes underwent a stabilization selection. qPCR based expression analysis showed diferent patterns of FaGMP paralogs during fruit ripening, while FaGMP4 expressed higher in the variety containing higher AsA. Overexpression of FaGMP4 in strawberry signifcantly enhanced AsA accumulation. Furthermore, the expression of FaGMP4 under the treatment of blue and red light was largely increased in leaves while signifcantly inhibited in fruit. These results revealed the vital role of FaGMP4 in regulating AsA in strawberry.

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        Evaluation of suitable qRT-PCR normalization genes for various citrus rootstocks

        He Wen,Xie Rui,Li Huan,Wang Yan,Chen Qing,Lin Yuanxiu,Zhang Yunting,Luo Ya,Zhang Yong,Tang Haoru,Wang Xiaorong 한국식물생명공학회 2022 Plant biotechnology reports Vol.16 No.1

        Citrus rootstock can modify plant growth and enhance stress resistance. There are many genotypes and species used as citrus rootstocks. Although multiple citrus rootstocks whole genome sequence and transcriptome databases have previously been published, no suitable internal reference genes have been investigated for standardization of gene expression via quantitative real-time PCR. Here we reported the first systematic and comprehensive analysis of reference genes for quantitative real- time PCR standardization in various citrus rootstocks. The expression stability of ten candidate reference genes in diverse sample subsets of flooding, drought, alkaline and cold treatments was evaluated using four statistical algorithms, geNorm, NormFinder, BestKeeper and ΔCt method. The results demonstrated that the expression stability of reference genes varied under different experimental conditions. In addition, normalization of gene expression of 9-cis-epoxycarotenoid dioxygenase 3 (NCED 3), involved in abscisic acid biosynthesis, was conducted to further confirm the reliability of the reference genes. Overall, EF-1α, DIM 1, GAPC and UBC expressed much more stably. ACTIN and GAPDH were not recommended for normalization in given experimental conditions due to low stability. Our main contribution was to identify reference genes with suitable and stable expression in citrus rootstocks varied across selected experimental conditions. Furthermore, these results will provide valuable information for future research on citrus rootstocks.

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