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Transgenic Wucai (Brassica campestris L.) produced via Agrobacterium-mediated anther transformation in planta

  • Guohu Chen
  • Fanli Zeng
  • Jian Wang
  • Xinyu Ye
  • Shidong Zhu
  • Lingyun Yuan
  • Jinfeng Hou
  • Chenggang WangEmail author
Original Article

Abstract

Key message

We developed a novel Agrobacterium-mediated anther transformation for Wucai in planta, and in this procedure, the male germ line was the predominant target.

Abstract

Wucai (Brassica campestris L.), a variant of non-heading Chinese cabbage, is widely cultured in China and only improved by classic breeding methods. Here, a novel and efficient in planta Agrobacterium-mediated anther transformation method is developed based on the optimization of several factors that affect anther transformation. After optimization, transformation with the manual pollination application led to increased transient GUS expression in anthers (reaching 91.59%) and the transformation efficacies in planta (0.59–1.56% for four commercial cultivars). The stable integration and inheritance of the transgenes were further examined by molecular and genetic analyses. Three T2 transgenic lines presented a segregation ratio of 3:1, which was consistent with the Mendelian feature of a single dominant gene. In addition, the GUS histochemical assay and genetic crossing analysis revealed that the male germ line was the predominant target in this transformation. This optimized transformation system could provide a useful tool for both the improvement of cultivar qualities and investigation of functional genes in Wucai.

Keywords

Anther-drop Cytological observation Genetic transformation In planta Non-heading Chinese cabbage 

Abbreviations

As

Acetosyringone

BAP

6-Benzyl amino purine

CCM

Co-cultivation medium

CDS

Coding sequence

CMS

Cytoplasmic male sterility

ESEM

Environmental scanning electron microscopy

MES

Morpholino ethane sulfonic acid

MS

Murashige and Skoog medium

GUS

β-Glucuronidase

Kan

Kanamycin

Rif

Rifampicin

SM

Selection medium

WT

Wild type

Notes

Acknowledgements

The National Natural Science Foundation of China (31801853); National Key R & D Program of China (2017YFD0101803); Provincial Science and Technology Major Project of Anhui (17030701013); and Major (KJ2017ZD15) and Key (KJ2017A153) Projects of Natural Science Research Funds in Support of Colleges supported this work.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no competing interests.

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Vegetable Genetics and Breeding Laboratory, College of HorticultureAnhui Agricultural UniversityHefeiChina
  2. 2.Anhui Provincial Engineering Laboratory of Horticultural Crop BreedingHefeiChina

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