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Missouri 1BL/1RS translocation stocks of Triticum turgidum (2n = 4x = Agric. Exp. Stn., Columbia. 28). Genome 36:1088–1091. Sears, R.G., T.S. Cox, and B.S. Gill. 1992. Registration of Hamlet, a Hessian fly resistant hard red winter wheat. Crop Sci. 32:506. Isolation and Characterization of Two Promoters from Linseed for Genetic Engineering Ravinder K. Jain,* R. Gail Thompson, David C. Taylor, Samuel L. MacKenzie, Alan McHughen, Gordon G. Rowland, Don Tenaschuk, and Maxine Coffey ABSTRACT manipulation. For example, there is a need for creating new linseed cultivars with a wider range of fatty acid Linseed (Linum usitatissimum L.) is an important oilseed crop composition to supplement the existing food and confec- worldwide and is cultivated for the high level of linolenic acid (18:3) in its seed oil. Currently, there is a concerted effort to improve linseed tions markets [17]. Also, there is commercial interest in by genetic engineering. This will require appropriate transgenes and the use of linseed as a vehicle for biofarming of pharma- tissue-specific or constitutive promoters. We report the isolation and ceutically related products because of its self-pollinating characterization of two linseed promoters from a two-member gene nature [13]. A need for linseed varieties tolerant to family encoding the enzyme stearoyl-acyl carrier protein desaturase various abiotic and biotic stresses has also been recog- (SAD). The SAD1 and SAD2 gene promoter were each fused tran- nized [17]. A number of herbicide-tolerant linseed varie- scriptionally with the reporter gene for -glucuronidase (uidA; GUS) ties that are useful in crop rotation programs are becom- and were transferred to linseed to study their expression pattern. In ing available [17]. transgenic linseed, GUS activity mediated by the SAD2 promoter Molecular genetic manipulation of linseed can be appeared to be constitutive and was detected in leaves, apices, stem, achieved by expressing appropriate transgenes by roots, flower buds, flowers, and seeds. In contrast, GUS activity medi- ated by the SAD1 promoter appeared to be root- and seed-specific. In means of tissue-specific or constitutive gene promoters. developing seeds, both the promoters exhibited a temporal expression A limited number of promoters have been used in lin- pattern concomitant with protein and lipid biosyntheses. The GUS seed to introduce novel characteristics. Whereas the activity could be detected as early as 4 days after pollination (dap) constitutive promoters such the CaMV 35S and nos gene and in mature seeds (50 dap) with the highest activities around mid- promoters have been shown to function in linseed [10], development. The first pair of linseed promoters will be useful for the seed specific napin promoter is not effective [14]. manipulating the expression of indigenous as well as transgenes in Moreover, these promoters are protected by intellectual linseed to create value-added cultivars. property laws, which cause unnecessary delays in reach- ing licensing agreements before the promoters can be used in a breeding program. We, therefore, set out to L inseed is the third most important oilseed crop in identify promoters from linseed that can be utilized in Canada and an important crop worldwide. It is our breeding programs. grown for the high linolenic acid (18:3) content in its In linseed, SAD (9-18:0-ACP desaturase; EC seed oil. There is a concerted effort by several labora- 1.14.99.6) activity can be detected from about 10 dap tories to diversify linseed as a crop by molecular genetic to seed maturity (R. Jain et al., 1996, unpublished), suggesting that the promoter of this gene would be use- ful in manipulating gene expression during seed devel- R.K. Jain, D.C. Taylor, S.L. MacKenzie, D. Tenaschuk, and M. Coffey, National Research Council of Canada, Plant Biotechnology Institute, opment. SAD is a soluble enzyme that catalyzes con- Saskatoon, SK, Canada S7N 0W9; R.G. Thompson, A. McHughen, version of stearoyl-ACP (18:0-ACP) to oleoyl-ACP and G.G. Rowland, Crop Development Centre, Univ. of Saskatche- (18:19-ACP) by introducing a double bond at the car- wan, Saskatoon, SK, Canada S7N 5A8. NRCC No. 42620 Received 2 Feb. 1999. *Corresponding author (Jravinder@pbi.nrc.ca). Abbreviations: dap, days after pollination; OLs, oligoribonucleotides; SAD, stearoyl-acyl carrier protein desaturase; uidA, -glucuronidase. Published in Crop Sci. 39:1696–1701 (1999). Published November, 1999