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Identification and functional analysis of a pollen fertility-associated gene GhGLP4 of Gossypium hirsutum L.

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Abstract

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Cotton male fertility-associated gene GhGLP4, encoding a germin-like protein, is essential for anthers development by keeping ROS homeostasis through reducing H2O2 level.

Abstract

Utilization of heterosis is an important way to increase cotton yield and improve fiber quality in hybrid cotton development programs. Male sterility is used in the development of cotton hybrids to reduce the cost of hybrid seed production by eliminating the process of emasculation. From the transcriptome analysis of genic male sterile mutant (ms1) and its background C312 of G. hirsutum, a gene encoding germin-like protein (GhGLP4) was found significantly down-regulated in different developmental stages of ms1 anthers. To explore the gene function in cotton fertility, GhGLP4 was further studied and interfered by virus-induced gene silencing. In the GhGLP4 interfered cotton lines, the expression level of GhGLP4 was significantly decreased in the stamens, and the down-regulation of GhGLP4 resulted in pollen sac closure, stigma exertion, filament shortening, decrease in the number of anthers and complete male sterility. The expression levels of respiratory burst oxidase homologs (Rboh, NADPH oxidase) were significantly altered. Further investigation showed that the SOD activity decreased while the H2O2 content increased in the atypical stamens. These results indicated that GhGLP4 gene affected the cotton anther development through maintenance of ROS homeostasis by H2O2 reduction.

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Abbreviations

MS:

Male sterility

ROS:

Reactive oxygen species

CMS:

Cytoplasmic male sterility

GMS:

Genic male sterility

PCD:

Programmed cell death

H2O2 :

Hydrogen peroxide

GLPs:

Germin-like proteins

SOD:

Superoxide dismutase

OXO:

Oxalate oxidase

AGPPase:

ADP glucose pyrophosphatase or phosphodiesterase

qRT-PCR:

Quantitative real-time PCR

PDS:

Phytoene desaturase

CTAB:

Cetyltriethylammnonium bromide

TTC:

2,3,5-Triphenylterazolium chloride

NBT:

Nitroblue tetrazolium chloride

ETC:

Mitochondrial electron transport chain

VIGS:

Virus-induced gene silencing

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Funding

This work was supported by National Natural Science Foundation of China (U1903204, 31671738). The funding agencies had no role in research design, data collection and analysis, or manuscript writing.

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Authors and Affiliations

  1. Plant Genomics and Molecular Improvement of Colored Fiber Laboratory, College of Life Sciences and Medicine, Zhejiang Sci-Tech University, Hangzhou, 310016, Zhejiang, China

    Hongli Zheng, Rongjia Wang , Qimeng Jiang , Diandian Zhang, Rongrong Mu, Yihan Xu, Vitalis E. Nnaemeka, Jun Mei, Yanyan Zhao, Fangfang Cai, Dongliang Yu, Yuqiang Sun & Liping Ke

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  1. Hongli Zheng
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Contributions

YS and LK conceived and designed the experiments. HZ, RW, QJ, DZ and LK performed the research and prepared the figures; RM, YX, VEN, JM, YZ, FC and DY participated in data analysis. LK, HZ, YS and DY wrote and corrected the article. All authors reviewed and approved the manuscript.

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Correspondence to Liping Ke.

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Zheng, H., Wang , R., Jiang , Q. et al. Identification and functional analysis of a pollen fertility-associated gene GhGLP4 of Gossypium hirsutum L.. Theor Appl Genet 134, 3237–3247 (2021). https://doi.org/10.1007/s00122-021-03888-x

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