Cluster Bud Induction of Soybean Whole Cotyledonary Node and Application in Agrobacterium mediated Genetic Transformation(PDF)

¡¶´ó¶¹¿Æѧ¡·[ISSN:1000-9841/CN:23-1227/S]

Issue:

2012Äê06ÆÚ

Page:

865-868

Research Field:

Publishing date:

Info

Title:

Cluster Bud Induction of Soybean Whole Cotyledonary Node and Application in Agrobacterium mediated Genetic Transformation

Author(s):

ZHANG Fu-li¹; SHU Wen-tao²; ZHANG Yi¹; GAO Dan¹; ZHANG Li¹; LU Ya-jie¹; LUO Xiang¹; LI Cheng-wei¹

1.Key Laboratory of Plant Genetics and Molecular Breeding,Department of Life Science,Zhoukou Normal University,Zhoukou 466001,Henan;

2.Zhoukou Academy of Agricultural Sciences,Zhoukou 466001,Henan,China

Keywords:

Soybean; Whole cotyledonary node; Regeneration; Agrobacterium mediated; Genetic transformation

PACS:

S565.1

DOI:

10.3969/j.issn.1000-9841.2012.06.001

Abstract:

The effects of nine concentration combinations of 6ª²BA,IBA and KT designed by orthogonal design[L9(3⁴)]were evaluated on induction of soybean[Glycine max(L.)Merrill\]multi shoot from whole cotyledonary nodes.Four soybean genotypes,including Zhouhuang 20,Zhoudou 02005,Zhoudou 11 and Zhoudou 18,were used as plant materials,while MSB was used as basic medium.The results showed that nine concentration combinations had different effects on induction of multi shoot for soybean whole cotyledonary nodes.The regeneration system was further used to Agrobacterium mediated genetic transformation.For nine concentration combinations,MSB+3.0 mg^.L^-16-BA+0.1 mg^.L^-1IBA+0.5 mg^.L^-1KT for Zhoudou 02005 and MSB+3.5 mg^.L^-16-BA+0.2 mg^.L^-1IBA+0.2 mg^.L^-1KT for Zhouhuang 20,Zhoudou 11 and Zhoudou18 were optimum to induce multi shoot than other combinations.For four genotypes,the regeneration efficiency of multi shoot for Zhouhuang 20 was the highest.The concentration of 0.8 mg^.L^-1 for IBA and 0.8-1.0 mg^.L^-1for NAA was much more suitable for soybean rooting,and rooting efficiency of Zhonghuang 20 was the highest. Agrobacterium mediated transformation of Zhonghuang 20 was performed using whole cotyledonary nodes as explants,it showed that reporter gene(GUS)had been transformed into soybean genome by PCR analysis and GUS staining with 7.8% transformation efficiency,and the period of obtaining transgenic plant was 35-42 d.

References:

[1]Hinchee M A W,Connor Ward D V,Newell C A,et al.Production of transgenic soybean plants using Agrobacterium mediated DNA transfer[J].Nature Biotechnology,1988,6:915-922.
[2]Liu S J,Wei Z M,Huang J Q.The effect of co-cultivation and selection parameters on Agrobacterium mediated transformation of Chinese soybean varieties[J].Plant Cell Reports,2008,27(3):489-498.
[3]Paz M M,Shou H,Guo Z,et al.Assessment of conditions affecting Agrobacterium mediated soybean transformation using the cotyledonary node explants[J].Euphytica,2004,136(2):167-179.
[4]Mello Farias P C,Chaves A L S.Advances in Agrobacterium mediated plant transformation with enphasys on soybean[J].Scientia Agricola,2008,65(1):95-106.
[5]Uranbey S,Sevimay C S,Ozcan S.Development of high frequency multiple shoot formation(Trifolium resupinatum L.)[J].Plant Cell,Tissue and Organ Culture,2005,80(2):229-232.
[6]Haliloglu K.Efficient regeneration system from wheat leaf base segments[J].Biologia Plantarum,2006,50(3):326-330.
[7]Ma X H,Wu T L.Rapid and efficient regeneration in soybean[Glycine max (L.)Merrill]from whole cotyledonary node explants[J].Acta Physiologiae Plantarum,2008,30(2):209-216.
[8]Cheng T Y,Saka H,Voqui Dinh T H.Plant regeneration from soybean cotyledonary node segments in culture[J].Plant Science Letters,1980,19(2):91-99.
[9]Murashige T,Skoog F.A revised medium for rapid growth and bioassays with tobacco tissue culture[J].Physiologia Plantarum,1962,15(3):473-497.
[10]Gamborg O L,Miller R A,Ojiama K.Nutrient requirements of suspension cultures of soybean root cells[J].Experimental Cell Research,1968,50(1):151-158.

[11]Olhoft P M,Flagel L E,Donovan C M,et al.Efficient soybean transformation using hygromycin B selection in the cotyledonary node method[J].Planta£¬2003,216(5):723-35.
[12]Jefferson R A,Kavanagh T A,Bevan M W.GUS fusions:glucuronidase as a sensitive and versatile gene fusion marker in higher plants[J].The EMBO Journal,1987,6(13):3901-3907.
[13]Kosugi S,Ohashi Y,Nakajima K,et al.An improved assay for glucuronidase in transformed cells:methanol almost completely suppresses a putative endogenous glucuronidase activity[J].Plant Science,1990,70(1):133-140.
[14]Somers D A,Samac D A,Olhoft P M.Recent advances in legume transformation[J].Plant Physiology,2003,131(3):892-899.
[15]Zhao Z Y,Gu W N,Cai T S,et al.High throughput genetic transformation mediated by Agrobacterium tumefaciens in maize[J].Molecular Breeding,2001,8(4):323-333.
[16]Jefferson R A.Assaying chimeric genes in plants:the GUS gene fusion system[J].Plant Molecular Biology Reporter,1987,5(4):387-405.
[17]Kagi C,Grob Hardt R.How females become complex:cell differentiation in the gametophyte[J].Current Opinion in Plant Biology,2007,10(6):633-638.

Memo

Memo:

-

Common functions