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
caliente

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
progeny ratio

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
(seeds per population)

Transgenic parent (seeds)

F1
(families)

F2
(families)

BC
(families)

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


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