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.
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1 2007, 34 (3) : Acta Horticulturae Sinica RAPD 13, 2, 1, 1 ( 1, ; 2, ) :,,, ;,, ;, F 1, , RAPD,, : ; ; ; ; ; RAPD : S : A: X (2007) 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 , China; 2 N ational Key Laboratory of Crop Genetics and Germ plasm Enhancem ent, N anjing Agricultural U niversity, N anjing , 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 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 : ; : : (BK ) 3 E2mail: mahx@ jaas1ac1cn
2 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; rm in m in ( 4) ; 9 ml 65,, 6530 m in; 15 ml (24 1),, rm in m in (15) ; 50 ml, 016,, 10 m in, rm in m in (RT) ; 70%, 5 ml TE DNA; 50L 5 mg L - 1 RNase, m in, (24 1),, rm in m in; 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, , ;,
3 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 Ellip se F. ananassashengxing Erect Few Few F Ellip se Ellip se F 1 F 1, 728, F 1 ( PMC) (2) PMC, , (1),, 9018%, 10%
4 F 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 % F. m andschurica (7) % F. ananassashengxing (0-2) (26-28) F % 9018% 411% 110% (4-10) (7-13) (1-4) (0-3) 1F F ig. 1D iak inesis of F 1 of F. m andschu rica F. ananassa A rrows show RAPD , bp(2, ),, DNA 12, 111DNA8 14, 103, 92%12, 106DNA, 12 10, S (2) 94, 8816% RAPD,
5 3 : RAPD 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. ananassashengxing S S S S S S S S S S S S : +, Note: +, show p resences of special bands from F. m andschurica, F. ananassashengxing, respectively.
6 S1402 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; Progenies from cross between F. m andschurica and F. ananassashengxing ; 13. Marker ( 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)
7 3 : RAPD 603 AAAπAπBBBB, F. vesca (AA ) ByrneJelenkovic (1976) F. nubicola,,, F. nubicola, F. nubicola, B ringhurst ( 1990 ), AAAπAπBBBπBπ, ( 2001) RAPD 13, ( F. orien talis),, 1, 1, 313RAPD RAPD(, 1997;, 1998) RAPD (, 2005) RAPD, DNA RAPD DNA (Hancock & Callow, 1994; Graham et al., 1996; Degani et al., 1998), 10,, RAPD, DNA,, References B ringhurst R S Cytogenetics and evolution in American Fragaria. HortScience, 25: Buuer A Progress in breeding decap loid Fragaria vescana. Acta Hortculturae, 348: Byrne D, Jelenkovic G Cytological diploidization in the cultivated octop loid strawberry Fragaria ananassa Can. J. Genet. Cytol., 18: Degani C L, Rowland J, LeviA, Hortynski J A, Galletta G J DNA fingerp rint of strawberry ( Fragaria ananassa) cultivars using random ly amp lified polymorphic DNA markers. Euphytica, 102: EvansW D. 1982a. Guelph SO 1 synthetic octop loid strawberry breeding clone. HortScience, 17: EvansW D. 1982b. Guelph SO 2 synthetic octop loid strawberry breeding clone. HortScience, 17: 834. Graham J, McN icol R J, McN icol J W A comparison ofmethods for the estimation of genetic diversity in strawberry cultivars. Theor. App l. Genet., 93: Hancock J F, Callow P A Random ly amp lified polymorphic DNA s in the cultivated strawberry, Fragaria ananassa. J. Amer. Sci., 119 (4) : Soc. Hort. Lei Jia2jun, Yang Gao, Dai Han2p ing, W u Lu2ping, Deng M ing2qin W ild strawberry resources in China. Journal of Fruit Science, 14
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