CRISPR/Cas9‐targeted mutagenesis of the BnaA03.BP gene confers semi‐dwarf and compact architecture to rapeseed ( Brassica napus L.)

作者：Shihang Fan, Liang Zhang, Min Tang, Ying Cai, Jinglin Liu, Hongfang Liu, Jing Liu, William Terzaghi, Hanzhong Wang, Wei Hua, Ming Zheng · 发表于：Plant Biotechnology Journal · 年份：2021 · DOI：10.1111/pbi.13703 · 被引用次数：55 · 研究领域：CRISPR and Genetic Engineering、Photosynthetic Processes and Mechanisms、Chromosomal and Genetic Variations

Plant height and branch angle are the key factors of rapeseed plant architecture that affect plant density and lodging, which are crucial for rapeseed yield. In China, ˜80% of growing areas are the semi-winter type (Bol-type), with taller plants that are easily affected by lodging and limit mechanical harvesting, especially of hybrids. Breeders have, therefore, been seeking an ‘ideotype’ for rapeseed breeding, deeming that semi-dwarf and compact architecture could benefit yield (Fu and Zhou, 2013; Li et al., 2019). However, no optimized plant architecture has been identified in rapeseed germplasm. In this study, we used the CRISPR/Cas9 gene editing system to knock out the BnaA03.BP gene [a homolog of Arabidopsis BREVIPEDICELLUS (BP)] creating a novel germplasm for optimizing rapeseed plant architecture. BP encodes a knotted1-like homeobox gene that plays a key role in the regulation of leaf morphogenesis and pedicel bending in Arabidopsis (Lincoln et al., 1994; Venglat et al., 2002). Blast analysis identified two close homologs of BP in the rapeseed genome (ZS11A03G024840 and ZS11C03G030900), which we named BnaA03.BP and BnaC03.BP. The two BnaBPs are highly conserved, sharing 87.5% and 86.5% identity with BP at the amino acid level respectively. In addition, the two BnaBP homologs share 98.1% identity in their nucleotide sequences with each other. RNA-seq and qRT-PCR analysis showed that transcripts of both BnaBPs were more abundant in wild type (WT) stems, and both genes dis...