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Transgenic Tobacco Expressing Zephyranthes grandiflora Agglutinin Confers Enhanced Resistance to Aphids

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Abstract

Plant lectins have been reported as transgenic resistance factors against a variety of insect pests. Herein, homologous analysis demonstrated that Zephyranthes grandiflora agglutinin (ZGA) exhibited high similarity with other monocot mannose-binding lectins (MBLs). Phylogenetic analysis revealed that it had taxonomical relationships with insecticidal MBLs. Subsequently, a plasmid expression vector pBI121 containing zga gene (pBIZGA) was constructed using the zga sequence, under the control of CaMV35S promoter and nos terminator. pBIZGA was then integrated into the genome of Nicotiana tabacum L. Polymerase chain reaction and Southern blot analysis demonstrated that this zga gene was integrated into the plant genome. Western blotting and agglutinating activity analysis also showed that transgenic tobacco plants expressed different levels of ZGA. Carbohydrate inhibition analysis indicated that recombinant ZGA and the native shared the same carbohydrate-binding specificity. Moreover, genetic analysis confirmed Mendelian segregation (3:1) of the transgenic in T1 progenies. In planta bioassays on T0 plants and their progenies indicated that expressed ZGA had an effect on reducing the survivability and fecundity of tobacco aphids (Myzus nicotianae). These findings demonstrate that the novel zga gene of ZGA can be expressed in crop plants susceptible to various sap-sucking insects.

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Abbreviations

pBIZGA:

expression vector pBI121 containing zga gene

ZGA:

Zephyranthes grandiflora agglutinin

CaMV35S:

35S cauliflower mosaic virus promoter

zga :

Zephyranthes grandiflora agglutinin gene

MBLs:

mannose-binding lectins

GNA:

Galanthus nivalis agglutinin

ZCA:

Zephyranthes candida agglutinin

gna :

Galanthus nivalis agglutinin gene

ASAL:

Allium sativum leaf agglutinin

WGA:

wheat germ agglutinin

6-BA:

6-benzylaminopurine

ConA:

concanavalin A

T0:

the first generation of transgenic plants

T1:

the first progenies of T0

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Acknowledgements

We thank Dr. Bo Liu and He-jiao Bian for their good comments on this manuscript. This work was supported in part by grants from the National Natural Science Foundation of China (General Programs: no. 30270331 and no. 30670469); Director Fund of State Key Laboratory of Oral Diseases (Sichuan University); The Science and Technology Fund for Distinguished Young Scholars of Sichuan Province (no. 06ZQ026-035); and the Key Technologies R&D Program of Sichuan Province (2006Z08-010).

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  1. College of Life Sciences, State Key Laboratory of Oral Diseases, Sichuan University, Chengdu, 610064, China

    Song-hua Ye, Sheng Chen, Fan Zhang, Wei Wang, Qin Tian, Jin-zhi Liu, Fang Chen & Jin-ku Bao

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  1. Song-hua Ye
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Correspondence to Jin-ku Bao.

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Ye and Chen contributed equally to this work.

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Ye, Sh., Chen, S., Zhang, F. et al. Transgenic Tobacco Expressing Zephyranthes grandiflora Agglutinin Confers Enhanced Resistance to Aphids. Appl Biochem Biotechnol 158, 615–630 (2009). https://doi.org/10.1007/s12010-008-8418-6

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