Alternative splicing of a barley gene results in an excess-tillering and semi-dwarf mutant
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An excess-tillering semi-dwarf gene Hvhtd was identified from an EMS-induced mutant in barley and alternative splicing results in excess-tillering semi-dwarf traits.
Tillering and plant height are important traits determining plant architecture and grain production in cereal crops. This study identified an excess-tillering semi-dwarf mutant (htd) from an EMS-treated barley population. Genetic analysis of the F1, F2, and F2:3 populations showed that a single recessive gene controlled the excess-tillering semi-dwarf in htd. Using BSR-Seq and gene mapping, the Hvhtd gene was delimited within a 1.8 Mb interval on chromosome 2HL. Alignment of the RNA-Seq data with the functional genes in the interval identified a gene HORVU2Hr1G098820 with alternative splicing between exon2 and exon3 in the mutant, due to a G to A single-nucleotide substitution at the exon and intron junction. An independent mutant with a similar phenotype confirmed the result, with alternative splicing between exon3 and exon4. In both cases, the alternative splicing resulted in a non-functional protein. And the gene HORVU2Hr1G098820 encodes a trypsin family protein and may be involved in the IAA signaling pathway and differs from the mechanism of Green Revolution genes in the gibberellic acid metabolic pathway.
We thank Professor Jerome Franckowiak (North Dakota State University) for providing the Bowman NILs. This work was supported by the Natural Science Foundation of Zhejiang Province (Grant No. Y20C130024), the National Natural Science Foundation of China (Grant No. 31571659, 31101149), the China Scholarship Council in 2017, National Key R&D Program of China (2018YFD0200500), the Key Research Foundation of Science and Technology Department of Zhejiang Province of China (2016C02050-9), China Agriculture Research System (CARS-05) and the GRDC (UMU00050).
Author contribution statement
WH and CL conceived the project; WH, CT, JHZ, YS, XQZ, JMW, XJW, JMY performed the research; WH, CT, JZX, and CL analyzed the data. WH and CL wrote the article with contributions from all authors.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
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