Validating Plant Genes Involved in Pepper Yellow Leaf Curl Indonesia Virus Infection Using VIGS in Model Plant Nicotiana benthamiana

Kusumanegara, Kusumawaty; Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Indonesian Agency for Agricultural Research and Development, Ministry of Agriculture, Jalan Tentara Pelajar 3A, Bogor 16111, West Java, Indonesia
Tel. +62-251-8337975; Fax. +62-251-8338820; Kaido, Masanori; Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan; Mise, Kazuyuki; Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan

Validating Plant Genes Involved in Pepper Yellow Leaf Curl Indonesia Virus Infection Using VIGS in Model Plant Nicotiana benthamiana

Files

Authors

Kusumanegara, Kusumawaty; Indonesian Center for Agricultural Biotechnology and Genetic Resources Research and Development, Indonesian Agency for Agricultural Research and Development, Ministry of Agriculture, Jalan Tentara Pelajar 3A, Bogor 16111, West Java, Indonesia Tel. +62-251-8337975; Fax. +62-251-8338820

Kaido, Masanori; Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan

Mise, Kazuyuki; Laboratory of Plant Pathology, Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto 606-8502, Japan

Publisher

Balai Besar Penelitian dan Pengembangan Bioteknologi dan Sumber Daya Genetik Pertanian

Description

Pepper yellow leaf curl disease caused by Pepper yellow leaf curl Indonesia virus (PepYLCIV) has become a challenge to chili pepper cultivation. Development of resistant variety by utilizing recessive resistance gene is expected to control the disease in the field. This study aimed to validate three plant genes, namely deltaCOP, hsc70, and BAM1, in PepYLCIV infection by applying Virus-induced Gene Silencing (VIGS) in a model plant, wild type Nicotiana benthamiana. PepYLCIV and construct of Tobacco rattle virus (TRV) which induced silencing of each gene were co-inoculated into N. benthamiana plants through agroinfiltration. Gene expression and the relative amount of viral DNA were determined by quantitative reverse transcription PCR (qRT-PCR) and quantitative PCR (qPCR), respectively, at 15 days post inoculation. The results showed a decreased level of deltaCOP, hsc70, and BAM1 expressions to 66.4%, 53.0%, and 47.0%, respectively, compared to that in the control (100%). Silencing of the three genes decreased the accumulation of PepYLCIV to 0.1%, 18.4%, and 63.0%, respectively, compared to that in the control. deltaCOP and hsc70 genes were indicated to be involved in the viral infection and could be good candidate genes for obtaining chili pepper varieties resistant to PepYLCIV. This result affirmed that the reverse genetics technique could be an alternative approach for identifying plant genes involved in viral infection, including PepYLCIV. The use of an infectious clone in this study allows the virus inoculations could be carried out without rearing and maintaining its natural vector, hence reduces the risk of virus transmission to healthy plants.

Keywords

PepYLCIV; VIGS; Nicotiana benthamiana; reverse genetics; quantitative PCR

URI

https://repository.pertanian.go.id/handle/123456789/10538

Collections

Jurnal AgroBiogen

Full item page