Ecological Archives C006-092-A1
Rogelio Cruz-Reyes, Germán Ávila-Sakar, Gumersindo Sánchez-Montoya, and Mauricio Quesada. 2015. Experimental assessment of gene flow between transgenic squash and a wild relative in the center of origin of cucurbits. Ecosphere 6:248. http://dx.doi.org/10.1890/es15-00304.1
Appendix A. Location of wild populations of gourds and additional details per crosses type and tested proportions, number of families per crosses in parental, hybrids, and backcrosses.
Table A1. Location of wild populations of C. argyrosperma sororia and distance (kilometers) between each.
Location |
Agua |
Camino a la Huerta |
Km 26.5 |
La culebra |
Limoncito |
Morelos |
Ranchitos |
Tecuan |
Tenacatita |
Agua caliente (19°21'12.52"N, 104°53'27.43"W) |
|
19.7 |
3.2 |
32.5 |
13.1 |
49.1 |
29.7 |
3.1 |
3.4 |
Camino a la Huerta (19°18'15.71"N, 104°42'45.72"W) |
|
|
16.2 |
15.9 |
28.8 |
65.6 |
45.3 |
20.8 |
17.9 |
Km 26.5 (19°20'25.34"N, 104°51'49.96"W) |
|
|
|
29.5 |
15.6 |
51.7 |
32.4 |
4.9 |
2.9 |
La culebra (19°11'54.72"N, 104°37'30.75"W) |
|
|
|
|
43.9 |
80.1 |
60.1 |
33.4 |
30.5 |
Limoncito (19°27'49.34"N, 104°56'8.25"W) |
|
|
|
|
|
36.3 |
16.86 |
14.7 |
16.4 |
Morelos (19°41'14.85"N, 105°11'17.59"W) |
|
|
|
|
|
|
20.6 |
48.9 |
51.5 |
Ranchitos (19°35'40.91"N, 105° 1'1.59"W) |
|
|
|
|
|
|
|
31.1 |
33.2 |
Tecuan (19°19'57.44"N, 104°54'36.64"W) |
|
|
|
|
|
|
|
|
3.0 |
Tenacatita (19º19`19.90¨ N, 104º 52`59.96¨W) |
|
|
|
|
|
|
|
|
|
Table A2. Pollen load treatments performed to obtain two generations of hybrids and first generation of backcrosses, derived from zucchini (C. pepo) transgenic resistant to viruses (VRT) and distant wild relative C. argyrosperma spp. sororia crosses.
Generation |
Treatment † |
Pollen recipient |
Pollen donor |
Type of cross |
Expected |
F1 |
Saturation |
Wild |
Transgenic |
VRT |
1:1 |
|
15:1 |
Wild |
Transgenic-Wild |
15:1 |
15:17 |
|
7:1 |
Wild |
Transgenic-Wild |
7:1 |
7:9 |
|
3:1 |
Wild |
Transgenic-Wild |
3:1 |
3:5 |
F2 |
Saturation |
F1 non transgenic hybrid |
F1 transgenic hybrid |
F1 non-VRT × F1 VRT |
1:1 |
|
Saturation |
F1 transgenic hybrid |
F1 transgenic hybrid |
F1 VRT × F1 VRT |
3:1 |
Backcross |
Saturation |
F1 transgenic hybrid |
Wild |
F1 VRT × W |
1:1 |
|
Saturation |
Wild |
F1 transgenic hybrid |
W × F1 VRT |
1:1 |
Notes;†For F1, the saturation treatment consisted on saturating the stigmata of wild plants using only transgenic pollen; ratios indicate mixed pollen loads in either a 15:1, 7:1, or a 3:1 transgenic:wild pollen ratio., F2 progeny were generated through two hand pollination treatments: (a) F1 non-VRT as pollen recipients and F1 VRT plants as pollen donors (F1 non-VRT × F1 VRT); (b) F1 VRT plants as both pollen donors and recipients (F1 VRT × F1 VRT). Backcrosses were generated using two hand pollination treatments: (a) F1 VRT as pollen recipients and wild plants as donors (F1 VRT × W); (b) wild recipients and F1 VRT hybrids as donors (W × F1 VRT). ‡Expected ratio of transgenic:non-transgenic progeny under Mendelian inheritance. Viability of pollen was checked in vivo once the pollinations were completed (approximately 20 minutes) for wild plants (0.86 proportion germination, n = 27 individuals), transgenic plants (0.88, n = 9), hybrids F1 (0.70, n = 13) and hybrids F2 (F1 non-VRT × F1 VRT; 0.75, n =9).
Table A3. Number of familes and seeds used per generation for crosses. In wild and transgenic parents we do not have data of kinship between plants.
Wild parent |
Transgenic parent (seeds) |
F1 |
F2 |
BC |
Agua caliente; 8, Camino a la huerta;8, km 26.5; 17 La culebra;11 Limoncito;7 Morelos;18 Ranchitos;11 El Tecuan; 14 Tenacatita; 15 |
50 |
Saturation; 40
15:1; 16 7:1; 12 3:1; 17 |
F1non-VRT × F1VRT; 27
F1VRT × F1VRT; 19 |
F1VRT × W; 4
W × F1VRT; 3 |