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Genetic and physical fine mapping of a multilocular gene Bjln1 in Brassica juncea to a 208-kb region

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Abstract

Most of the germplasm resources in Brassica juncea produce silique with only two locules, whereas a few varieties can produce silique with three or four locules. The increase in locule number in B. juncea has been shown to cause an increase in the number of seeds per silique, resulting in an increase in the yield per plant. Thus, the development of high-locule-number varieties may be an effective way of improving the yield of B. juncea. Duoshi, a B. juncea landrace originating from the Qinghai–Tibetan plateau, produces silique with 3–4 locules. Genetic analysis has shown that the high-locule-number trait in Duoshi is determined by two recessive genes, tentatively designated as Bjln1 and Bjln2. For fine mapping of the Bjln1 gene, a BC3 population was developed from the cross between Duoshi (multilocular parent) and Xinjie (bilocular parent). Using a combination of amplified fragment length polymorphism (AFLP) and bulked segregant analysis, only two AFLP markers linked to Bjln1 were identified. Preliminary linkage analysis showed that the two AFLP markers were located on the same side of Bjln1. Blast analysis revealed that the sequences of the two AFLP markers had homologues on Scaffold000019 at the bottom of B. rapa A7. Using the results of linkage analysis and BlastN searches, simple sequence repeat (SSR) markers were subsequently developed based on the sequence information from B. rapa A7. Seven SSR markers were eventually identified, of which ln 8 was co-segregated with Bjln1. ln 7 and ln 9, the closest flanking markers, were mapped at 2.0 and 0.4 cM distant from the Bjln1 gene, respectively. The SSR markers were cloned, sequenced and mapped on A7 of B. rapa (corresponding to J7 in the A genome of B. juncea). The two closest flanking markers, ln 7 and ln 9, were mapped within a 208-kb genomic region on B. rapa A7, in which the Bjln1 gene might be included. The present study may facilitate cloning of the Bjln1 gene as well as the selection process for developing multilocular varieties in B. juncea by marker-assisted selection and genetic engineering.

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Acknowledgments

The authors are grateful to Dr. Xinhua Zeng, Xiaoling Dun and Shengqian Xia for technical assistance. This research was financially supported by funds from the national nature science funds of China (31060196), the High-tech program “863” (2011AA10A104), the National Key Basic Research Program of China “973” (2012CB723007) and the science and technology program of Qinghai province (2010-Z-707).

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Authors and Affiliations

  1. Qinghai Academy of Agricultrual and Forestry Sciences, Key Laboratory of Spring Rape Genetic Improvement of Qinghai Province, National Key Laboratory Breeding Base for Innovation and Utilization of Plateau Crop Germplasm, Xining, 810016, Qinghai, China

    Lu Xiao, Huiyan Zhao, Zhi Zhao, Dezhi Du, Liang Xu, Yanmei Yao, Zhigang Zhao, Xiaorong Xing, Guoxia Shang & Hongchao Zhao

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  1. Lu Xiao
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  2. Huiyan Zhao
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  3. Zhi Zhao
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  4. Dezhi Du
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  5. Liang Xu
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  6. Yanmei Yao
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  7. Zhigang Zhao
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  8. Xiaorong Xing
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  9. Guoxia Shang
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  10. Hongchao Zhao
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Correspondence to Dezhi Du.

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Supplementary Figure 1

Schematic representation of the pedigree of the genetic stocks and the population used for mapping of the Bjln1 gene (jpg 75 kb)

Supplementary Figure 2

The SSR amplification results from 20 ml and 20 bl plants. The arrows indicate the polymorphic bands present in bl individuals but not in ml individuals. The SSR markers used was ln 5 (a) and ln 8 (b), respectively. M 100-bp DNA ladder (jpg 34 kb)

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Xiao, L., Zhao, H., Zhao, Z. et al. Genetic and physical fine mapping of a multilocular gene Bjln1 in Brassica juncea to a 208-kb region. Mol Breeding 32, 373–383 (2013). https://doi.org/10.1007/s11032-013-9877-1

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