Identification and evaluation of the novel genes for transcript normalization during female gametophyte development in sugarcane

Maokai Yan; Xingyue Jin; Yanhui Liu; Huihuang Chen; Tao Ye; Zhimin Hou; Zhenxia Su; Yingzhi Chen; Mohammad Aslam; Yuan Qin; Xiaoping Niu

doi:10.7717/peerj.12298

Identification and evaluation of the novel genes for transcript normalization during female gametophyte development in sugarcane

PeerJ. 2021 Oct 19:9:e12298. doi: 10.7717/peerj.12298. eCollection 2021.

Authors

Maokai Yan^#¹, Xingyue Jin^#², Yanhui Liu², Huihuang Chen², Tao Ye², Zhimin Hou², Zhenxia Su², Yingzhi Chen¹, Mohammad Aslam¹, Yuan Qin^{1

2}, Xiaoping Niu¹

Affiliations

¹ Guangxi Key Laboratory of Sugarcane Biology, State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources, College of Agriculture, Guangxi University, Nanning, China.
² Key Lab of Genetics, Breeding and Multiple Utilization of Crops, Ministry of Education, Fujian Provincial Key Laboratory of Haixia Applied Plant Systems Biology, College of Life Sciences, Fuzhou, China.

^# Contributed equally.

Abstract

Background: Sugarcane (Saccharum spontaneum L.), the major sugar and biofuel feedstock crop, is cultivated mainly by vegetative propagation worldwide due to the infertility of female reproductive organs resulting in the reduction of quality and output of sugar. Deciphering the gene expression profile during ovule development will improve our understanding of the complications underlying sexual reproduction in sugarcane. Optimal reference genes are essential for elucidating the expression pattern of a given gene by quantitative real-time PCR (qRT-PCR).

Method: In this study, based on transcriptome data obtained from sugarcane ovule, eighteen candidate reference genes were identified, cloned, and their expression levels were evaluated across five developmental stages ovule (AC, MMC, Meiosis, Mitosis, and Mature).

Results: Our results indicated that FAB2 and MOR1 were the most stably expressed genes during sugarcane female gametophyte development. Moreover, two genes, cell cycle-related genes REC8 and CDK, were selected, and their feasibility was validated. This study provides important insights into the female gametophyte development of sugarcane and reports novel reference genes for gene expression research on sugarcane sexual reproduction.

Keywords: Female gametophyte; Novel genes; Sugarcane; Transcriptome analysis.

Grants and funding

This work was supported by the Science and Technology Major Project of Guangxi (Gui Ke 2018-266-Z01), the National Natural Science Foundation of China (31800262; U1605212; 31761130074; 31600249; 31700279), the China Postdoctoral Science Foundation (2018M632564), The Weng Hongwu Academic Innovation Research Fund of Peking University, and a Guangxi Distinguished Experts Fellowship. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.