Knockout of miR396 genes increases seed size and yield in soybean

Hongtao Xie ¹Fei Su ²Qingfeng Niu ³Leping Geng ³Xuesong Cao ⁴Minglei Song ⁴Jinsong Dong ²Zai Zheng ⁵Rui Guo ²Yang Zhang ²Yuanwei Deng ³Zhanbo Ji ⁶Kang Pang ⁶Jian-Kang Zhu ⁴Jianhua Zhu³

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作者信息

1. School of Life Sciences,Anhui Agricultural University,Hefei 230036,China;Research Center for Biological Breeding Technology,Research Institute of Frontier Science,Anhui Agricultural University,Hefei 230036,China;Bellagen Biotechnology Co.Ltd,Jinan 250000,China
2. Institute of Crop Sciences/National Nanfan Research Institute,Chinese Academy of Agricultural Sciences,Key Laboratory of Gene Editing Technologies(Hainan),Ministry of Agriculture and Rural Affairs,Sanya 572025,China
3. School of Life Sciences,Anhui Agricultural University,Hefei 230036,China;Research Center for Biological Breeding Technology,Research Institute of Frontier Science,Anhui Agricultural University,Hefei 230036,China
4. Southern University of Science,and Technology,Shenzhen 518055,China
5. Hainan Yazhou Bay Seed Laboratory,Sanya 572024,China
6. Bellagen Biotechnology Co.Ltd,Jinan 250000,China
折叠

Abstract

Yield improvement has long been an important task for soybean breeding in the world in order to meet the increasing demand for food and animal feed.miR396 genes have been shown to negatively regulate grain size in rice,but whether miR396 family members may function in a similar manner in soybean is un-known.Here,we generated eight soybean mu-tants harboring different combinations of ho-mozygous mutations in the six soybean miR396 genes through genome editing with clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated nuclease(Cas)12SF01 in the elite soybean cultivar Zhong-huang 302(ZH302).Four triple mutants(mir396aci,mir396acd,mir396adf,and mir396cdf),two quadruple mutants(mir396-abcd and mir396acfi),and two quintuple mu-tants(mir396abcdf and mir396bcdfi)were characterized.We found that plants of all the mir396 mutants produced larger seeds com-pared to ZH302 plants.Field tests showed that mir396adf and mir396cdf plants have sig-nificantly increased yield in growth zones with relatively high latitude which are suited for ZH302 and moderately increased yield in lower latitude.In contrast,mir396abcdf and mir396bcdfi plants have increased plant height and decreased yield in growth zones with rela-tively high latitude due to lodging issues,but they are suited for low latitude growth zones with increased yield without lodging problems.Taken together,our study demonstrated that loss-of-function of miR396 genes leads to sig-nificantly enlarged seed size and increased yield in soybean,providing valuable germ-plasms for breeding high-yield soybean.

Key words

Cas12SF01/CRISPR/Cas/miR396/seed size/soy-bean/yield

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基金项目

Bellagen Biotechnology Inc.,and by the National Key Research and Development Program of China(NK2022010301)

Seed-Industrialized Development Program in Shandong Province,China(2021LZGC003)

Key Research & Development Program of Shandong Province,China(2021LZGC012)

Innovative research team of Anhui Education(2022AH010056)

National Natural science Foundation of China(32270367)

PhD Scientific Research and Innovation Foundation of Sanya Yazhou Bay Science and Technology City(HSPHDSRF-2023-11-007)

出版年

2024

植物学报(英文版)

中国植物学会

植物学报(英文版)

CSTPCDCSCD

影响因子：0.921

ISSN：1672-9072

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