Genome-wide identification of new reference genes for RT-qPCR normalization in CGMMV-infected Lagenaria siceraria

PeerJ

article

Chenhua Zhang¹  Hongying Zheng²  Xinyang Wu²  Heng Xu³  Kelei Han²  Jiejun Peng²  Yuwen Lu²  Lin Lin²  Pei Xu²  Xiaohua Wu²  Guojing Li²  Jianping Chen¹  Fei Yan² 

[1] College of Life Sciences, Fujian Agriculture and Forestry University;The State Key Laboratory Breeding Base for Sustainable Control of Pest and Disease, Zhejiang Academy of Agricultural Sciences;Key Laboratory of Biotechnology in Plant Protection of MOA of China and Zhejiang Province, Institute of Virology and Biotechnology, Zhejiang Academy of Agricultural Sciences;Institute of Vegetable, Zhejiang Academy of Agricultural Sciences

关键词: RT-qPCR;    Reference gene;    Transcriptome;    Bottle gourd;    CGMMV;   

DOI  :  10.7717/peerj.5642

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

Lagenaria siceraria is an economically important cucurbitaceous crop, but suitable reference genes (RGs) to use when the plants are infected by cucumber green mottle mosaic virus (CGMMV) have not been determined. Sixteen candidate RGs of both leaf and fruit and 18 candidate RGs mostly from separate RNA-Seq datasets of bottle gourd leaf or fruit were screened and assessed by RT-qPCR. The expression stability of these genes was determined and ranked using geNorm, NormFinder, BestKeeper and RefFinder. Comprehensive analysis resulted in the selection of LsCYP, LsH3, and LsTBP as the optimal RGs for bottle gourd leaves, and LsP4H, LsADP, and LsTBP for fruits. LsWD, LsGAPDH, and LsH3 were optimal for use in both leaves and fruits under the infection of CGMMV. Isopentenyl transferase (IPT) and DNA-directed RNA polymerase (DdRP) were used to validate the applicability of the most stable identified RGs from bottle gourd in response to CGMMV. All the candidate RGs performed in RT-qPCR consistently with the data from the transcriptome database. The results demonstrated that LsWD, LsGAPDH and LsH3 were the most suitable internal RGs for the leaf, and LsH3, LsGAPDH, LsP4H and LsCYP for the fruit.

【 授权许可】

CC BY