Agrobacterium tumefaciens-mediated transformation of a hevein-like gene into asparagus leads to stem wilt resistance.

Helong Chen,Jianming Gao,Jian Wang,Shiqing Zhang,Jingen Xi,Kexian Yi,Jinlong Zheng,Xing Huang,Shibei Tan,Zhiwei Lu,Anping Guo

doi:10.1371/journal.pone.0223331

Abstract

Asparagus stem wilt, is a significant and devastating disease, typically leading to extensive economic losses in the asparagus industry. To obtain transgenic plants resistant to stem wilt, the hevein-like gene, providing broad spectrum bacterial resistance was inserted into the asparagus genome through Agrobacterium tumefaciens-mediated transformation. The optimal genetic transformation system for asparagus was as follows: pre-culture of embryos for 2 days, inoculation using a bacterial titre of OD600 = 0.6, infection time 10 min and co-culturing for 4 days using an Acetosyringone concentration of 200 μmol/L. Highest transformation frequencies reached 21% and ten transgenic asparagus seedlings carrying the hevein-like gene were identified by polymerase chain reaction. Moreover, integration of the hevein-like gene in the T1 generation of transgenic plants was confirmed by southern blot hybridization. Analysis showed that resistance to stem wilt was enhanced significantly in the transgenic plants, in comparison to non- transgenic plants. The results provide additional data for genetic improvement and are of importance for the development of new disease-resistant asparagus varieties.

Highlights

Asparagus (Asparagus officinalis L.), known as the “King of Vegetables”, is among the most popular nutrient-rich and healthy vegetables because of its unique texture, taste and high nutrient content [1]
XbaI and SacI restriction sites were added to both ends of the hevein-like gene amplification product, and the PCR amplification product was subjected to DNA digestion
The large fragment of plasmid pBI121 and the hevein-like gene were ligated and the products transformed into E.Coli DH5α

Summary

Introduction

Asparagus (Asparagus officinalis L.), known as the “King of Vegetables”, is among the most popular nutrient-rich and healthy vegetables because of its unique texture, taste and high nutrient content [1]. Asparagus has various bioactive properties [2]. Research on development of improved Asparagus germplasm in China is lacking. Stem wilt (Phoma asponaqi Sacc) of asparagus is a severe disease that leads to huge economic losses. Transgenic breeding of Asparagus has not been exploited and only three reports regarding.

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Results

Conclusion