2007, 34 (3) : 597-604 Acta Horticulturae Sinica RAPD 13, 2, 1, 1 ( 1, 210014; 2, 210095) :,,, ;,, ;, F 1, 611+ 1016+ 211+ 0135, RAPD,, : ; ; ; ; ; RAPD : S 66814 : A: 05132353X (2007) 0320597208 Cytogenetics and RAPD Ana lysis of In terspec if ic Hybr ids from the Cross of F raga ria m andschu rica Staudt and F. ananassa D uch. MA Hong2xiang 13, CHEN Pei2du 2, YU Gui2hong 1, and REN L i2juan 1 ( 1 Institute of A gro2biological Genetics and Physiology, J iangsu A cadem y of A gricultural Sciences, N anjing 210014, China; 2 N ational Key Laboratory of Crop Genetics and Germ plasm Enhancem ent, N anjing Agricultural U niversity, N anjing 210095, China) Abstract: The interspecific hybrid was obtained from the cross of F raga ria m andschu rica Staudt, dip loid species derived from Northeast China, and F. ananassa Duch. (2n = 8x = 56) Shengxingby using immature embryo culture technique. The hybrids were pentap loids w ith poor fertility. The morphological char2 acteristics of the hybrids were observed and showed that they were intermediate between their parents in pedun2 cle height and p lant height, and sim ilar to F raga ria m andschu rica in hair on peduncle and petiole. The hybrids showed expected 35 chromosomes in somatic cells and had a meiotic chromosome association of 611 + 1016 + 211 + 0135 with a few abnormal m icrospores occurring in tetrad phase. RAPD analysis showed that the hybrids possessed the specific bands of two parents, and the genetic diversities were found among the interspecific hybrid lines. Key words: Strawberry; F raga ria m andschu rica Staudt; F raga ria ananassa Duch. ; Interspecific hybridization; Cytogenetics; RAPD a,,, ( Evans, 1982a, 1982b; Buu2 : 2006-10 - 27; : 2007-03 - 06 : (BK2001417) 3 E2mail: mahx@ jaas1ac1cn
598 34 er, 1993; Trajkvoski, 1993, 1997; Sangiacomo & Sullivan, 1994; Yuji, et al, 1997) 10 (,, 2002, 2004;, 2004) 1997), ( ( F. m andschu rica), ( 2001 ),, (, 2002), (, 2003),,, RAPD, 1, 1999,,,, (Carnoy), 45%, ( PMC) DNA CTAB, 4 g, 20 ml, 50 ml,, 10 m in; 3 750 rm in - 1 20 m in ( 4) ; 9 ml 65,, 6530 m in; 15 ml (24 1),, 3 750 rm in - 1 20 m in (15) ; 50 ml, 016,, 10 m in, 2 500 rm in - 1 10 m in (RT) ; 70%, 5 ml TE DNA; 50L 5 mg L - 1 RNase, 371030 m in, (24 1),, 10 000 rm in - 1 10 m in; 011 3 moll - 1 (ph 512) DNA, 70% DNA, TE RAPD Sangon RAPD 25 L, 30 ngdna, 1 buffer, 4 dntp 012 mmoll - 1, 2 mmoll - 1 MgCl 2, 20 ng, 110 U TaqDNA( ) PCR PE29600 : 941 m in; 941 m in, 382 m in, 722 m in45; 725 m in 5L, 20L 118%,, 1LmL - 1 EB40 m in, 2 211 1, ;,
3 : RAPD 599,, ;, ;, 14 (2n = 2x) 56 (2n = 8x), F 1 35 (2n = 5x) 1F 1 Table 1M orpholog ic characters of F 1 from the cross of F. m andschu rica F. ananassa Materials Choromoso2 me number (2n) Plant height ( cm) Plant form Leaf shape Hair on petiole Stamen Peduncle Hair on peduncle F. m andschurica 14 1617 Ellip se 56 2910 F. ananassashengxing Erect Few Few F 1 1921 35 2419 1922 35 2812 1923 35 3010 1924 35 2614 1925 35 2515 Ellip se 1926 35 2618 1927 35 2417 1928 35 2212 Ellip se 1929 35 2616 19210 35 2818 212F 1 F 1, 728, F 1 ( PMC) 6110+ 10160+ 2110+ 0135(2) PMC, 101606110, (1),, 9018%, 10%
600 34 2F 1 Table 2Chrom osom e conf igra tion of PM C a t m etapha seand ra tio of d ifferen t k inds of m icrospore in hybrids from F. m andschu rica F. ananassa Materials Number of cell observed Univalent B ivalent Trivalent Tetravalent Number of m icrospore 3 4 5 6 10 7100 100% F. m andschurica (7) 21 0186 27157 100% F. ananassashengxing (0-2) (26-28) F 1 20 6110 10160 2110 0135 411% 9018% 411% 110% (4-10) (7-13) (1-4) (0-3) 1F 1 51021 F ig. 1D iak inesis of F 1 of F. m andschu rica F. ananassa A rrows show 5+ 10 + 2 + 1. 213RAPD 312 12 18, 26 700 bp(2, ),, DNA 12, 111DNA8 14, 103, 92%12, 106DNA, 12 10, S1402 10(2) 94, 8816% RAPD,
3 : RAPD 601 312 Table 3Spec ia l bands from paren ts and 10 offspr ings from F. m andschu rica F. ananassashengx ingusing 12 pr im ers Primers DNA bands F. m andschurica F 1 F. ananassashengxing1921 1922 1923 1924 1925 1926 1927 1928 1929 19210 S1364 2-3 + - + + + 5 + - + + + + + + + + S1365 1 2 4 + - + + + + + + + 8 + - + + S1371 6 7 11 S1376 1 3 + + + + + + + + 7 + + + + + + + + S1380 1 + + + + + + 2 5 8 S1397 1 3 + + + 5 7 + + + + + + + + + S1399 2 3 6 + + + + + + + + 7 S1402 2 3 4 6 + + + 7 + + + S1404 4 5 8 + + + + + + + + S1405 3 8 9 11 S1409 3 6 + + + + + + + 9 + + + + + + + + + + + 13 + + + + + + S1414 1 3 + + + + + + + 7 + + + + + + + + : +, Note: +, show p resences of special bands from F. m andschurica, F. ananassashengxing, respectively.
602 34 2S1402 RAPD 1. ; 2. ; 312. ; 13. F ig. 2RAPD bands of the parents and their progen ies from F. m andschu rica and F. ananassa Shengx ingby primer S1402 Arrow shows a special amplified band of Shengxing. 1. F. m andschurica; 2. Shengxing; 3-12. Progenies from cross between F. m andschurica and F. ananassashengxing ; 13. Marker. 3 311 ( Sjulin & Dale, 1987), Evans ( 1982a, 1982b) Buuer (1993) Trajkvoski (1993, 1997) Yuji( 1997),,, (, 1997),,, (2002, 2004) 4x5x6x7x8x9x (, 2004),,,,, 312 ( F. virginiana Duch. ) ( F. chiloensis Duch. ), SenanayakeB ringhurst (1967)
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