Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (1): 1-13.doi: 10.3864/j.issn.0578-1752.2016.01.001

• CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS •     Next Articles

Observation, Genetic Analysis and Gene Mapping of an Open Hull Semi-Sterility Mutant in Rice (Oryza sativa)

CHEN Li-kai, HUANG Ming, LIU Yong-zhu, WANG Hui, CHEN Zhi-qiang, GUO Tao   

  1. National Engineering Research Centre of Plant Space Breeding, South China Agricultural University, Guangzhou 510642
  • Received:2015-07-13 Online:2016-01-01 Published:2016-01-01

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Abstract: 【Objective】The paper is order to carry out a morphological characteristics investigation and genetic analysis of an open-hull semi-sterility mutant (ohss) of rice (Oryza sativa L.), induced by spaceflight, laid the groundwork for further gene cloning and function analysis by fine mapping and preliminary screening of the candidate gene responsible for the mutated trait.【Method】The ohss mutant was derived from rice variety Hanghui 7, which was induced on Spaceship “Shenzhou 8”. The morphological characteristics of ohss were anatomically observed to analyze the mutagenic features of floral organ development. Subsequently, pollen fertility, natural seed setting rate and bagged seed setting rate were investigated for fertility evaluation. Five plants of ohss mutation and WT were random selected to survey the panicle and grain related traits. SSR markers covering the whole genome were used to detect the mutagenic effect of ohss. Moreover, genetic analysis was conducted using the crosses between ohss and three wild type varieties, Hanghui 7, Francis and 02428 where the phenotypes of the F1 and F2 were surveyed and c2 test was performed. A population from the cross of 02428/ohss was used to map the ohss(t) geneusing SSR markers and newly developed InDel markers. The candidate gene was predicted based on the RAP gene annotation database of the mapping regionand screened through sequences alignment and expression of candidate genes.【Result】Compared with wild-type, panicles of mutant ohss were enclosed and florets showed abnormalities at the reproductive stage, and the palea and lemma were weak, distorted and dehiscent, with organ similar to the palea in the floret, while some of florets had no palea differentiation. Sterility testing showed that pollen grain rate of the abnormal spikelet of ohss was 58.7%, leading to significantly lower seed set, less panicle weight per plant and filled grain number per panicle compared with the wild-type. Mutation survey based on SSR markers revealeda total of 0.0336 of variation frequency was caused between ohss and WT, and variation frequency of different chromosome varied from 0.0143 to 0.0889, except chromosome 7 and 12. Genetic analysis indicated that the mutant phenotype in ohss was controlled by a single recessive nuclear gene, namely ohss(t), which had been fine mapped to a 27.6 kb physical distance between two InDel markers, InDel6043 and InDel6070 on chromosome 3, where three annotated genes were predicted. Based on the result of sequencing, semi-quantitative RT-PCR and real time quantitative PCR, there was no mutation occurring in the coding and promoter sequence of OsMADS1 of ohss,but strong changes on gene expression pattern.【Conclusion】The mutated trait of ohss was controlled by a single recessive nuclear gene ohss(t), which was fine mapped to a 27.6 kb physical distance between InDel6043 and InDel6070 on chromosome 3. No nucleotide sequence mutation was found to occur in the coding sequence or the 5′UTR of OsMADS1,but expression of OsMADS1 was strongly inhibited in ohss(t).

Key words: rice (Oryza sativa L.), spaceflight mutagenesis, floral organ development, genetic analysis, gene mapping

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