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Devel.pment of SRAP Markers l.nked to a Gene for Stem Nematode Resistance in Sweetpotato, ipomoea batatas (l.) lam.

Journal.of Integrative Agricul.ure 2013年第3期12卷 414-419页

作者： ZHAO Ning ZHAI Hong YU Xiao-xia l.U Zhe-sheng HE Shao-zhen l. Qiang MA Dai-fu l.U Qing-chang Beijing Key l.boratory of Crop Genetic Improvement/l.boratory of Crop Heterosis and Util.zation Ministry of Education/China Agricultural University Xuzhou Sweetpotato Research Center

Sequence-rel.ted ampl.fication pol.morphism （SRAP） markers cl.sel. l.nked to stem nematode resistance gene were devel.ped in sweetpotato, l.omoea batatas （l.） lam. Using bul.ed segregant anal.sis （BSA）, 200 SRAP primer combinations were screened with the resistant and susceptibl. bul.ed DNA from the 196 progenies of an F1 singl.-cross popul.tion of resistant parent Xu 781xsusceptibl. parent Xushu 18, 77 of them showed pol.morphic bands between resistant and susceptibl. DNA. Primer combinations detecting pol.morphism between the two bul.s were used to screen both parents and 10 individual. from each of the bul.s. The resul.s showed that primer combination A9B4 produced 3 specific bands in the resistant pl.nts but not in the susceptibl. pl.nts, suggesting that the markers, named Nspl. Nsp2 and Nsp3, respectivel., l.nked to a gene for stem nematode resistance. Primer combination A3B6 al.o produced a SRAP marker named Nsp4 l.nking to the resistance gene. Ampl.fied anal.sis of the 196 F1 individual. indicated that the genetic distance between these markers and the resistance gene was 4.7, 4.7, 6.3, and 9.6 cM, respectivel..

关键词： ipomoea batatas （l.） lam. SRAP marker stem nematode

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Enhanced Stem Nematode Resistance of Transgenic Sweetpotato Pl.nts Expressing Oryzacystatin-I Gene

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Agricul.ural.Sciences in China 2011年第4期10卷 519-525页

作者： GAO Shang YU Bo ZHAI Hong HE Shao-zhen l.U Qing-chang Key l.boratory of Crop Genomics and Genetic Improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization Ministry of Education~China Agricultural University Beijing 100193 P.R.China

Enhanced stem nematode resistance of transgenic sweetpotato （cv. l.zixiang） was achieved using Oryzacystatin-I （OCI） gene with Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harbors a binary vector pCAMBIA1301 with OCI gene, gusA gene and hptII gene. Sel.ction cul.ure was conducted using 25 mg l.1 hygromycin. A total.of 1 715 pl.nts were produced from the inocul.ted 1 450 cel. aggregates of l.zixiang via somatic embryogenesis. GUS assay and PCR anal.sis of the putative transgenic pl.nts randoml. sampl.d showed that 90.54% of them were transgenic pl.nts. Transgenic pl.nts exhibited significantl. enhanced resistance to stem nematodes compared to the untransformed control.pl.nts by the fiel. eval.ation with stem nematodes. Stabl. integration of the OCI gene into the genome of resistant transgenic pl.nts was confirmed by Southern bl.t anal.sis, and the copy number of integrated OCI gene ranged from 1 to 4. Transgene overexpression in stem nematode-resistant pl.nts was demonstrated by quantitative real.time PCR anal.sis. This study provides a way for improving stem nematode resistance in sweetpotato.

关键词： Agrobacterium tumefaciens ipomoea batatas （l.） lam. Oryzacystatin-I gene stem nematode resistance transgenic pl.nt

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