Resistance to Fusarium oxysporum f. sp. gladioli in transgenic Gladiolus plants expressing either a bacterial chloroperoxidase or fungal chitinase genes

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• 作者：Kathryn Kamo ; Dilip Lakshman ; Ruchi Pandey…
• 关键词：Floral monocots ; Stable transformation ; Fungus resistance
• 刊名：Plant Cell, Tissue and Organ Culture
• 出版年：2016
• 出版时间：March 2016
• 年：2016
• 卷：124
• 期：3
• 页码：541-553
• 全文大小：1,539 KB
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• 作者单位：Kathryn Kamo (1)
  Dilip Lakshman (1)
  Ruchi Pandey (1)
  Mary Ann Guaragna (1)
  Patricia Okubara (2)
  Kanniah Rajasekaran (3)
  Jeffrey Cary (3)
  Ramon Jordan (1)
  
  1. Floral and Nursery Plants Research Unit, USDA, National Arboretum, Beltsville, MD, USA
  2. Root Disease and Biological Control Research Unit, USDA, Pullman, WA, USA
  3. Food and Feed Safety Research Unit, Southern Regional Research Center, USDA, New Orleans, LA, USA
  
• 刊物类别：Biomedical and Life Sciences
• 刊物主题：Life Sciences
  Plant Sciences
  Plant Physiology
  
• 出版者：Springer Netherlands
• ISSN：1573-5044

文摘

Three antifungal genes, a non-heme chloroperoxidase from Pseudomonas pyrrocinia, and an exochitinase and endochitinase from Fusarium venetanum under regulation by the CaMV 35S promoter, were used to transform Gladiolus for resistance to Fusarium oxysporum f. sp. gladioli. Gladiolus plants were confirmed to be transgenic by Southern hybridization. Semi-quantitative RT-PCR of RNA isolated from leaves and roots demonstrated expression of the Fusarium exochitinase and endochitinase genes in transgenic plants compared to controls. All transgenic plants expressing the Fusarium exochitinase or endochitinase gene had chitinase activity higher than that of the control plants. Semi-quantitative RT-PCR verified that three of the four plant lines with the chloroperoxidase gene expressed the transgene in leaves and roots while no expression was detected in control plants. Western hybridization confirmed the presence of the chloroperoxidase protein in both leaves and roots of transgenic plants. Cell extracts from one endochitinase plant line inhibited growth of germinated F. oxysporum spores more consistently than extracts from the four chloroperoxidase and three endochitinase plant lines. Three chloroperoxidase, two exochitinase, and three endochitinase transgenic plant lines sustained a significantly (P < 0.05) lower density of hyphae on roots compared to roots of non-transformed Gladiolus plants three to four days following exposure of the roots to Fusarium. Shoots from two plant lines, one containing a chloroperoxidase and the other an endochitinase gene, had less necrosis when rated on a scale of 1–3 and appeared visually to be healthier and without obvious Fusarium infection than non-transformed, regenerated Gladiolus plants 17–21 days following exposure to F. oxysporum. Keywords Floral monocots Stable transformation Fungus resistance