Abstract

Rice is one of the staple crops in the world. Grain size is an important determinant of rice grain yield, but the genetic basis of the grain size remains unclear. Here, we report a set of chromosome segment substitution lines (CSSL) developed in the genetic background of the genome-sequenced indica cultivar Zhenshan 97. Genotyping of the CSSLs by single nucleotide polymorphism array shows that most carry only one or two segments introduced from the genome-sequenced japonica cultivar Nipponbare. Using this population and the high-density markers, a total of 43 quantitative trait loci were identified for seven panicle- and grain-related traits. Among these loci, the novel locus qGL11 for grain length and thousand-grain weight was validated in a CSSL-derived segregating population and finely mapped to a 25-kb region that contains an IAA-amido synthetase gene OsGH3.13, This gene exhibited a significant expression difference in the young panicle between the near-isogenic lines that carry the contrasting Zhenshan 97 and Nipponbare alleles at qGL11. Expression and sequence analyses suggest that this gene is the most likely candidate for qGL11. Furthermore, several OsGH3.13 mutants induced by a CRISPR/Cas9 approach in either japonica or indica exhibit an increased grain length and thousand-grain weight, thus enhancing the final grain yield per plant. These findings provide insights into the genetic basis of grain size for the improvement of yield potential in rice breeding programs.

Highlights

  • Rice (Oryza sativa L.) is the staple food for more than half of the world’s population

  • It has been reported that chromosome segment substitution lines (CSSL) as an advanced backcross population can effectively reduce the interaction between quantitative trait loci (QTL) and minimize the genetic background interference, improving the power to detect, fine-map and clone Q­ TLs31–36

  • Most CSSLs carry multiple and long substituted chromosomal segments with a low marker density, which could decrease the detection power, especially for those minor QTLs that are much more masked by genetic interactions

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Summary

Introduction

Rice (Oryza sativa L.) is the staple food for more than half of the world’s population. QTGW3, a major QTL for grain weight, encodes a SHAGGY-like kinase 41 (OsSK41)/OsGSK5, which interacts with auxin response factor 4 (OsARF4) to negatively regulate grain size and grain weight by affecting cell ­expansion[5]. BIG GRAIN1 (BG1) encodes a positive regulator of auxin response and transport, which markedly increased the indole-3-acetic acid (IAA) level in the panicle. A large number of CSSL populations have been developed with great effort in rice and other crops. Using the developed CSSLs with high-density SNP markers, we performed QTL analysis for seven panicle and grainrelated traits and identified a number of QTLs with minor effects on the assayed traits. The CRISPR-generated OsGH3.13 mutants exhibited increased grain length and grain weight, indicating that the regulation of grain size involves the IAA metabolic pathway

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