Constitutive expression of a cowpea trypsin inhibitor gene, CpTi, in transgenic rice plants confers resistance to two major rice insect pests
Tóm tắt
The gene encoding a cowpea trypsin inhibitor (CpTI), which confers insect resistance in trangenic tobacco, was introduced into rice. Expression of the CpTi gene driven by the constitutively active promoter of the rice actin 1 gene (Act1) leads to high-level accumulation of the CpTI protein in transgenic rice plants. Protein extracts from transgenic rice plants exhibit a strong inhibitory activity against bovine trypsin, suggesting that the proteinase inhibitor produced in transgenic rice is functionally active. Small-scale field tests showed that the transgenic rice plants expressing the CpTi gene had significantly increased resistance to two species of rice stem borers, which are major rice insect pests. Our results suggest that the cowpea trypsin inhibitor may be useful for the control of rice insect pests.
Tài liệu tham khảo
Boulter D, Gatehouse AMR, Hilder V: Use of cowpea trypsin inhibitor (CpTI) to protect plants against insect predation. Biotechnol Adv 7: 489–497 (1989).
Cao J, Duan X, McElroy D, Wu R: Regeneration of herbicide resistant transgenic rice plants following micro-projectile-mediated transformation of suspension culture cells. Plant Cell Rep 11: 586–591 (1992).
Chrispeels MJ, Raikhel NV: Lectins, lectin genes and their role in plant defense. Plant Cell 3: 1–19 (1991).
Fujimoto H, Itoh K, Yamamoto M, Kyozuka J, Shimamoto K: Insect resistant rice generated by introduction of a modified δ-endotoxin gene of Bacillus thuringiensis. Bio/technology 11: 1151–1155 (1993).
Gasser CS, Fraley RT: Transgenic crops. Sci Am 266: 62–70 (1992).
Hayakawa T, Zhu Y, Itoh K, Kimura Y, Izawa T, Simamoto K, Toriyama S: Genetically engineered rice resistant to rice stripe virus, an insect-transmitted virus. Proc Natl Acad Sci USA 89: 9865–9869 (1992).
Heinrichs EA, Medrano FG, Rapusas HR: Genetic Evaluation for Insect Resistance in Rice. International Rice Research Institute, Los Baños, Philippines (1985).
Hilder VA, Gatehouse AMR, Sheerman SE, Barker RF, Boulter D: A novel mechanism of insect resistance engineered into tobacco. Nature 330: 160–163 (1987).
Hilder VA, Barker RF, Samour RA, Gatehouse AMR, Gatehouse JA, Boulter D: Protein and cDNA sequences of Bowman-Birk protease inhibitors from the cowpea (Vigna unguiculata Walp.). Plant Mol Biol 13: 701–710 (1989).
Hoffmann MP, Zalon FG, Wilson LT, Smilanick JM, Malyj LD, Kiser J, Hilder VA, Barnes WM: Field evaluation of transgenic tobacco containing genes encoding Bacillus thuringiensis δ-endotoxin or cowpea trypsin inhibitor: efficiency against Helicoverpa zea (Lepidoptera: Noctuidae). J Econ Entomol 85: 2516–2522 (1992).
Huesing JE, Shade RE, Chrispeels MJ, Murdock LL: α-Amylase inhibitor, not phytohemagglutinins explains the resistance of common bean seeds to cowpea weevil. Plant Physiol 96: 993–996 (1991).
Johnson R, Narvaez J, An G, Ryan C: Expression of proteinase inhibitors I and II in transgenic tobacco plants: Effects on natural defense against Manduca sexta larvae. Proc Natl Acad Sci USA 86: 9871–9875 (1989).
Kao KN: Chromosomal behavior in somatic hybrids of soybean — Nicotiana glauca. Mol Gen Genet 150: 225–230 (1977).
Ling KC: Rice Virus Diseases, pp. 11–33. International Rice Research Institute, Los Baños, Philippines (1972).
Masoud SA, Johnson LB, White FF, Reeck GR: Expression of a cysteine proteinase inhibitor (oryzacystatin-I) in transgenic tobacco plants. Plant Mol Biol 21: 655–663 (1993).
McElroy D, Blowers A, Jenes B, Wu R: Construction of expression vectors based on the rice actin 1 (Act1) 5′ region for use in monocot transformation. Mol Gen Genet 231: 150–160 (1991).
McElroy D, Zhang W, Wu R: Isolation of an efficient actin promoter for use in rice transformation. Plant Cell 2: 163–171 (1990).
Murashige T, Skoog F: A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15: 473–497 (1962).
Ryan CA: Protease inhibitors in plants: Genes for improving defense against insects and pathogens. Annu Rev Phytopath 28: 425–449 (1990).
Ryan CA: Proteinase inhibitor gene families: Strategies for transformation to improve plant defense against herbivores. BioEssays 10: 20–24 (1989).
Schroeder HE, Gollasch S, Moore A, Tabe LM, Graig S, Hardie DC, Chrispeels MJ, Spencer D, Higgins TJV: Bean α-amylase inhibitor confers resistance to the pea weevil (Bruchus pisorum) in transgenic peas (Pisum sativum L.). Plant Physiol 107: 1233–1239 (1995).
Schwert GW, Takenaka Y: A spectrophotometric determination of trypsin and chymotrypsin. Biochim Biophys Acta 16: 570–575 (1955).
Shade RE, Schroeder HE, Pueyo JJ, Tabe LM, Murdock LL, Higgins TJV, Chrispeels MJ: Transgenic pea seeds expressing the alpha-amylase inhibitor of the common bean are resistant to bruchid beetles. Bio/technology 12: 793–796 (1994).
Wolfson JL, Murdock LL: Diversity in digestive proteinase activity among insects. J Chem Ecol 16: 1089–1102 (1990).
Wu R, Duan X, Xu D: Analysis of rice genes in transgenic plants. Progr Nucl Acid Res Mol Biol 45: 1–26 (1993).
Xu D, Wu T, Cao J, Wu R: Production and analysis of transgenic rice plants. In: You CB, Chen ZL, Ding Y (eds) Biotechnology in Agriculture, pp. 130–135. Kluwer Academic Publishers, Dordrecht, Netherlands (1993).
Zhang W, McElroy D, Wu R: Analysis of rice Act 5′ region activity in transgenic rice plants. Plant Cell 3: 1155–1165 (1991).
Zhang W, Wu R: Efficient regeneration of transgenic rice plants from rice protoplasts and correctly regulated expression of foreign genes in the plants. Theor Appl Genet 76: 835–840 (1988).