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Morphogenesis and Gene Mapping of deformed interior floral organ 1 (difo1), a Novel Mutant Associated with Floral Organ Development in Rice

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Abstract

Floral organ identity and specific number directly affect anthesis habits, fertilization and grain yield. Here, we identified a deformed interior floral organ 1 (difo1) mutant from selfing progenies of indica cv. Zhonghui8015 (Zh8015) after 60Co γ-ray treatment. Compared with the Zh8015 spikelet, the interior floral organs of the difo1 mutant present various numbers of stamens and stigmas, with no typical filament and no mature pollen grains. Most difo1 flowers exhibited an increased number of stigmas that were attached to the stamens and an intumescent ovule-like cell mass in addition to the ovary. Transverse sections of spikelets and scanning electron microscopy analysis revealed an indeterminate number of interior floral organs and abnormal early spikelet development for the difo1 mutant. Instead of the linear-shaped surface of wild-type stamens, difo1 displayed a glossy stamen surface resulting in immature stamens and complete sterility. In addition, the difo1 mutant exhibited delayed anthesis, rapid anthesis and non-extended stamens compared with wild type. Genetic analysis and gene mapping revealed that difo1 was controlled by a single recessive gene, which was fine-mapped to a 54-kb interval on the short arm of chromosome 4 between markers S22 and RM16439 harboring nine ORFs. Sequence analysis revealed that the mutant carried a single nucleotide deletion in its promoter region, which likely corresponded to the phenotype, in a C2H2-type zinc finger protein gene (LOC_Os04g08600). Moreover, qRT-PCR analysis showed a significantly down-regulated expression pattern for DIFO1 and many floral organ identity genes in the interior floral organs of difo1. DIFO1 is therefore an important floral organ development gene in rice, particularly with regard to interior organ meristem identity and floret primordium differentiation.

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Abbreviations

CTAB:

Cetyltrimethyl ammonium bromide

InDel:

Insertion/deletion

kb:

Kilobase

ORF:

Open reading frame

PCR:

Polymerase chain reaction

qRT-PCR:

Quantitative real-time PCR

SEM:

Scanning electron microscope

SSR:

Simple sequence repeats

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Acknowledgments

We thank Professor Dabing Zhang and Wanqi Liang (Shanghai Jiao Tong University) for their kind guidance in the analysis of semi-thin sections. This work was supported by the National Special Fund for Agro-scientific Research in the Public Interest (Grants 20140302 and 20150308), the National Key Transform Program (Grants 2014ZX08001-002), the National Natural Science Foundation of China (Grants 31501290), the Zhejiang Provincial Natural Science Foundation of China (Grants Q13C130009), and the Super Rice Breeding Innovation Team and Rice Heterosis Mechanism Research Innovation Team of the Chinese Academy of Agricultural Sciences Innovation Project (Grants CAAS-ASTIP-2013-CNRRI).

Author Contributions

Shihua Cheng, Liyong Cao and Yingxin Zhang conceived and designed the study. Lianping Sun conducted the experiments andreceived help from Peipei Zhang, Zhengfu Yang, Xiaoxiao Zhou, Dandan Xuan and Zihe Li in the sampling and cytological observations. Weixun Wu, Xiaodeng Zhan, Xihong Shen, Ping Yu and Daibo Chen guided and participated in the anthesis habit analysis. Lianping Sun and Peipei Zhang collected and analyzed the data. Lianping Sun, Yingxin Zhang and Peipei Zhang wrote the paper. Lianping Sun, Yingxin Zhang, Peipei Zhang, Zhengfu Yang, Xiaoxiao Zhou, Dandan Xuan, Zihe Li, Weixun Wu, Xiaodeng Zhan, Xihong Shen, Ping Yu, Daibo Chen, Li-yong Cao and Shi-hua Cheng reviewed and edited the manuscript. All authors read and approved the manuscript.

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    Authors and Affiliations

    1. College of Agronomy, Shenyang Agricultural University, Shenyang, Liaoning, 110866, China

      Lianping P. Sun, Xiaoxiao X. Zhou & Shihua H. Cheng

    2. National Center for Rice Improvement/China National Rice Research Institute/ Key Laboratory for Zhejiang Super Rice Research, Hangzhou, Zhejiang, 31140, China

      Lianping P. Sun, Yingxin X. Zhang, Peipei P. Zhang, Zhengfu F. Yang, Dandan D. Xuan, Md Habibur H. Rahman, Zihe H. Li, Weixun X. Wu, Xiaodeng D. Zhan, Xihong H. Shen, Ping Yu, Daibo B. Chen, Liyong Y. Cao & Shihua H. Cheng

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    Lianping P. Sun, Yingxin X. Zhang and Peipei P. Zhang contributed equally to this work.

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    Sun, L.P., Zhang, Y.X., Zhang, P.P. et al. Morphogenesis and Gene Mapping of deformed interior floral organ 1 (difo1), a Novel Mutant Associated with Floral Organ Development in Rice. Plant Mol Biol Rep 35, 130–144 (2017). https://doi.org/10.1007/s11105-016-1007-x

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