Rapid Identification of Pollen- and Anther-Specific Genes in Response to High-Temperature Stress Based on Transcriptome Profiling Analysis in Cotton

Abstract

Anther indehiscence and pollen sterility caused by high temperature (HT) stress have become a major problem that decreases the yield of cotton. Pollen- and anther-specific genes play a critical role in the process of male reproduction and the response to HT stress. In order to identify pollen-specific genes that respond to HT stress, a comparative transcriptome profiling analysis was performed in the pollen and anthers of Gossypium hirsutum HT-sensitive Line H05 against other tissue types under normal temperature (NT) conditions, and the analysis of a differentially expressed gene was conducted in the pollen of H05 under NT and HT conditions. In total, we identified 1111 pollen-specific genes (PSGs), 1066 anther-specific genes (ASGs), and 833 pollen differentially expressed genes (DEGs). Moreover, we found that the late stage of anther included more anther- and pollen-specific genes (APSGs). Stress-related cis-regulatory elements (CREs) and hormone-responsive CREs are enriched in the promoters of APSGs, suggesting that APSGs may respond to HT stress. However, 833 pollen DEGs had only 10 common genes with 1111 PSGs, indicating that PSGs are mainly involved in the processes of pollen development and do not respond to HT stress. Promoters of these 10 common genes are enriched for stress-related CREs and MeJA-responsive CREs, suggesting that these 10 common genes are involved in the process of pollen development while responding to HT stress. This study provides a pathway for rapidly identifying cotton pollen-specific genes that respond to HT stress.

A male-sterile mutant with necrosis-like dark spots on anthers was generated in cotton

“… Creation of Gh EMS1 genes mutants by using CRISPR/Cas9. (A) A phylogenetic tree for EMS1 genes in Gossypium hirsutum and AtEMS1 was constructed using the neighbor-joining method in MEGA 10.1.8 followed by bootstrapping with 1,000 replicates; (B) Expression profiles of GhEMS1 genes in flower buds of different lengths (3~5 mm, 5~6 mm, 6~7 mm, 7~8 mm, 8~9 mm, 9~10 mm, 10~11 mm, 11~12 mm, 12~13 mm, 13~14 mm, 14~16 mm, 16~19 mm, 19~24 mm)in G. hirsutum H05 determined by transcriptome sequencing ( Zhang et al., 2022 ); (C) Creation of a male-sterile line pool by CRISPR/Cas9. Two sgRNAs were serially connected to each vector to increase the knockout efficiency; (D) Two pairs of sgRNAs on exons were designed and vectors containing polycistronic tRNA–gRNA genes ( PTG ).…”

High‐temperature stress in crops: male sterility, yield loss and potential remedy approaches

“…HT (Zhang et al, 2022b). Even a single day HT stress will affect the development stages of flowers, the reproductive process (Zinn et al, 2010), the anther structure and microspore cellular morphology (Hedhly et al, 2009).…”

“…Plant Biotechnology Journal published by Society for Experimental Biology and The Association of Applied Biologists and John Wiley & Sons Ltd., 21, 680-697fertility during HT by using ZFNs and TALENs are few as compared to those using the clustered regularly interspaced short palindromic repeats (CRISPR)/Cas system. In the recent study ofZhang et al (2022b), anther specific genes were studied in cotton during HT stress. Among these genes, the authors obtained the genome edited line (Ghir_D12G012350), and the gene functions in anther dehiscence and affects fertility during HT stress.…”

Response of root and root hair phenotypes of cotton seedlings under high temperature revealed with RhizoPot

“…Temperature has an extremely significant effect on cotton growth and development, and it can affect its growth and maturation and regulate its phenological development and rate of biomass accumulation ( Xu et al., 2017 ). Temperature beyond a certain range (35°C) seriously affects the germination and elongation of pollen tubes ( Zahid et al., 2016 ), causes indehiscent anthers and sterile pollen, and reduces the single boll weight, which ultimately leads to a decrease in crop yield ( Zhang et al., 2022 ). Each time the daily maximum temperature rises by 1°C, the lint yield will decrease by 110 kg/hm 2 ( Gao et al., 2021 ).…”