We compared some fitness components of the wild squash species Cucurbita pepo spp. ovifera var. texana (C. texana) and three generations of hybrids (F1, BC1, and BC2) between C. texana and commercial transgenic squash CZW-3 over three consecutive years under field conditions of low (LDP) and high disease pressure (HDP) by Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV) and Watermelon mosaic virus (WMV). Transgenic squash CZW-3 expresses the coat protein (CP) genes of CMV, ZYMV, and WMV, and is resistant to these three aphid-borne viruses. Across all HDP trials, transgenic BC1 and BC2 hybrids expressing the three CP genes grew more vigorously, displayed resistance to CMV, ZYMV, and WMV, and produced a greater number of mature fruits and viable seeds than nontransgenic hybrid segregants and C. texana. Transgenic F1 hybrids behaved similarly to BC1 and BC2 hybrids but grew less vigorously than C. texana. In contrast, across all LDP trials, C. texana outperformed the transgenic and nontransgenic hybrid segregants. Further, only one back cross was necessary to recover individuals with most of the C. texana characteristics and yet maintain virus resistance. Our data suggest that C. texana acquiring CP transgenes upon hybridization and introgression could have a selective advantage if CMV, ZYMV, and WMV are severely limiting the growth and reproductibility of wild squash populations.

We monitored pollen-mediated transgene dissemination from commercial transgenic squash CZW-3 into its wild relative Cucurbita pepo ssp. ovifera var. texana (C. texana). Transgenic squash CZW-3 expresses the neomycin phosphotransferase II (nptII) gene and the coat protein (CP) genes of Cucumber mosaic virus (CMV), Zucchini yellow mosaic virus (ZYMV), and Watermelon mosaic virus (WMV); thereby, it is resistant to these three aphid-borne viruses. The rate of NPT II and CP transgene introgression increased with overlapping flowering patterns and a high ratio of transgenic F1 hybrids (C. texana × CZW-3) to C. texana. Transgene transfer also readily occurred from transgenic F1 hybrids into C. texana over three generations in field settings where test plants grew sympatrically and viruses were not severely limiting the growth, and fruit and seed production of C. texana. In contrast, introgression of the transgenes into C. texana was not sustained under conditions of high viral disease pressure. As expected, C. texana progeny that acquired the CP transgenes exhibited resistance to CMV, ZYMV, and WMV. This is the first report on transgene dissemination from a transgenic crop that exhibits disease resistance and hybridizes with a wild plant species without loss of fertility.

The effect of preplant conditioning on germination of three flower seeds, Bupleurum griffithii (Tourn.) L. (thorough-wax), Ammi majus L. (greater ammi), and Cirsium japonicum DC. Per. (Japanese thistle), were studied. Seeds were osmoconditioned with -1.2 MPa polyethylene glycol 8000 (PEG) solution and matriconditioned with moist Micro-Cel E (ratio of 2 seed: 0.6 carrier: 3 water by weight for Bupleurum and Cirsium; for Ammi the ratio was 2:1.4:6) and moist expanded vermiculite #5 (the ratio was of 2 seed: 0.6 carrier: 2 water for Bupleurum). In some treatments, water in the matriconditioning mixture was replaced with 1 mm gibberellin A4+7 (GA) or 0.2 % KNO3. In Bupleurum, matriconditioning with Micro-Cel E was generally superior to matriconditioning with vermiculite or osmoconditioning with PEG. A 4-day matriconditioning with Micro-Cel E and germination in the dark reduced the period required for 50% (T50 of final germination by 4 days and improved the percentage germination at 20C (73 % vs. 95%), compared to nonconditioned seeds germinated in the dark. The treatment also improved the percentage of germination at 15C (68% vs. 95%) and effectively removed the thermoinhibition of germination at 25 and 30C. Germination was inhibited to a greater extent for seeds kept in the light during matriconditioning and germination than for seeds conditioned in darkness and germinated in light or conditioned in light and germinated in darkness. Nitrate added during conditioning in light prevented inhibition of germination, provided seeds were kept in darkness during germination. In A. majus, germination in light after 4-day matriconditioning reduced the T50 by ≈2 days, but had little effect on percentage germination. Both GA and irradiance equally promoted germination when added during osmoconditioning, with nitrate having no effect. In C. japonicum, a 4-day matriconditioning or a 7-day osmoconditioning reduced the T50 of germination by -2 days and improved the percentage germination to some extent. Neither irradiance nor nitrate had any significant effect.