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2013 | 35 | 01 |
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Vacuum infiltration enhances the Agrobacterium-mediated genetic transformation in Indian soybean cultivars

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For the first time we have developed a reliable and efficient vacuum infiltration-assisted Agrobacteriummediated genetic transformation (VIAAT) protocol for Indian soybean cultivars and recovered fertile transgenic soybean plants through somatic embryogenesis. Immature cotyledons were used as an explant and three Agrobacterium tumefaciens strains (EHA 101, EHA 105, and KYRT 1) harbouring the binary vector pCAMBIA1301 were experimented in the co-cultivation. The immature cotyledons were pre-cultured in liquid somatic embryo induction medium prior to vacuum infiltration with the Agrobacterium suspension and co-cultivated for 3 days on co-cultivation medium containing 50 mg l-1 citric acid, 100 lM acetosyringone, and 100 mg l-1 L-cysteine. The transformed somatic embryos were selected in liquid somatic embryo induction medium containing 10 mg l-1 hygromycin and the embryos were germinated in basal medium containing 20 mg l-1 hygromycin. The presence and integration of the hpt II and gus genes into the soybean genome were confirmed by GUS histochemical assay, polymerase chain reaction, and Southern hybridization. Among the different combinations tested, high transformation efficiency (9.45 %) was achieved when immature cotyledons of cv. Pusa 16 were pre-cultured for 18 h and vacuum infiltrated with Agrobacterium tumefaciens KYRT 1 for 2 min at 750 mm of Hg. Among six Indian soybean cultivars tested, Pusa 16 showed highest transformation efficiency of 9.45 %. The transformation efficiency of this method (VIAAT) was higher than previously reported sonication-assisted Agrobacterium-mediated transformation. These results suggest that an efficient Agrobacteriummediated transformation protocol for stable integration of foreign genes into soybean has been developed.
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  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Department of Biotechnology, School of Life Sciences, Bharathidasan University, Tiruchirappalli 620024, Tamil Nadu, India
  • Acereto-Escoffie´ POM, Chi-Manzanero BH, Echeverria-Echeverria S, James A, Grijalva R, Rodriguez-Zapata LC (2005) Agrobacterium-mediated transformation of Musa acuminata cv ‘‘Grand Nain’’ scalps by vacuum infiltration. Sci Hort 105:359–371
  • Amoah BK, Wu H, Sparks C, Jones HD (2001) Factors influencing Agrobacterium-mediated transient expression of uidA in wheat inflorescence tissue. J Exp Bot 52:1135–1142
  • An G (1985) High-efficiency transformation of cultured tobacco cells. Plant Physiol 79:568–570
  • Bailey MA, Boerma HR, Parrott WA (1993) Genotype effects on proliferative embryogenesis and plant regeneration of soybean. In Vitro Cell Dev Biol Plant 29P:102–108
  • Barwale UB, Kerns HR, Widholm JM (1986a) Plant regeneration from callus cultures of several soybean genotypes via embryogenesis and organogenesis. Planta 167:473–481
  • Barwale UB, Meyer MM, Widholm JM (1986b) Screening of Glycine max and Glycine soja genotypes for multiple shoot formation at the cotyledonary node. Theor Appl Genet 72:423–428
  • Bechtold N, Pelletier G (1998) In-Planta Agrobacterium-mediated transformation of adult Arabidopsis thaliana plants by vacuum infiltration. Methods Mol Biol 82:259–326
  • Bechtold N, Ellis J, Pelletier G (1993) In-Planta Agrobacteriummediated gene transfer by infiltration of adult Arabidopsis thaliana plants. CR Acad Sci 316:1194–1199
  • Birt DF, Hendrich S, Anthony M, Alkel DL (2004) Soybeans and the prevention of chronic human disease. In: Specht J, Boerma R (eds) Soybeans: improvement, production and uses, 3rd edn. American society of Agronomy, Madison, pp 1047–1117
  • Canche-Moo RLR, Ku-Gonzalez A, Burgeff C, Loyola-Vargas VM, Rodriguez-Zapata LC, Castano E (2006) Genetic transformation of Coffea canephora by vacuum infiltration. Plant Cell Tissue Org Cult 84:373–377
  • Cervera M, Pina JA, Juarez J, Navarro L, Pena L (1998) Agrobacterium-mediated transformation of citrange: factors affecting transformation and regeneration. Plant Cell Rep 18:271–278
  • Charity JA, Holland L, Donaldson SS, Grace L, Walter C (2002) Agrobacterium-mediated transformation of Pinus radiata organogenic tissue using vacuum infiltration. Plant Cell Tissue Org Cult 70:51–60
  • Cheng TY, Saka H, Voqui-Dinh TH (1980) Plant regeneration from soybean cotyledonary node segments in culture. Plant Sci Lett 19:91–99
  • Christou P, McCable DE, Martinell BJ, Swain WF (1990) Soybean genetic engineering-commercial production of transgenic plants. Trends Biotechnol 8:145–151
  • Curtis IS, Power JB, Hedden P, Ward DA, Phillips A, Lower KC, Davey MR (1999) A stable transformation system for the ornamental plant, Datura meteloides (D.C.). Plant Cell Rep 18:554–560
  • Dan Y, Reichert NA (1998) Organogenic regeneration of soybean from hypocotyl explants. In Vitro Cell Dev Biol Plant 34:14–21
  • Dellaporta SL, Wood J, Hicks JB (1983) A plant DNA mini preparation: Version II. Plant Mol Biol Rep 1:19–21
  • Dong JJ, Kharb P, Teng W, Hall TC (2001) Characterization of rice transformed via an Agrobacterium-mediated inflorescence approach. Mol Breed 7:187–194
  • Finer JJ, McMullen MD (1991) Transformation of soybean via particle bombardment of embryogenic suspension cultures. In Vitro Cell Dev Biol Plant 27P:175–182
  • Finer JJ, Nagasawa A (1988) Development of an embryogenic suspension culture of soybean (Glycine max Merrill.). Plant Cell Tissue Org Cult 15:125–136
  • Franklin G, Carpenter L, Davis E, Reddy CS, Al-Abed D, Alaiwi WA, Parani M, Smith B, Goldman SL, Sairam RV (2004) Factors influencing regeneration of soybean from mature and immature cotyledons. Plant Growth Regul 43:73–79
  • Gamborg OL, Miller RA, Ojima K (1968) Nutrient requirements of suspension cultures of soybean root cells. Exp Cell Res 50:150–158
  • Gupta S, Gupta S, Bhat V, Gupta MG (2006) Somatic embryogenesis and Agrobacterium-mediated genetic transformation in Indian accessions of Lucerne (Medicago sativa L.). IJBT 5:269–275
  • Hardegger M, Sturm A (1998) Transformation and regeneration of carrot (Daucus carota L.). Mol Breed 4:119–127
  • Hiei Y, Komari T, Ishida Y, Saito H (2000) Development of Agrobacterium-mediated transformation method for monocotyledonous plants. Breed Res 2:205–213
  • Hinchee M, Connor-Ward DV, Newell CA, McDonnell RE, Sato SJ, Gasser CS, Fischhoff DA, Re DB, Fraley RT, Horsch RB (1988) Production of transgenic soybean plants using Agrobacteriummediated DNA transfer. Nat Biotechnol 6:915–922
  • Hong HP, Zhang H, Olhoft P, Hill S, Wiley H, Toren E, Hillebrand H, Jones T, Cheng M (2007) Organogenic callus as the target for plant regeneration and transformation via Agrobacterium in soybean (Glycine max (L.) Merr.). In Vitro Cell Dev Biol Plant 43:558–568
  • Hood EE, Helmer GC, Fraley RT, Chilton MD (1986) The hypervirulence of Agrobacterium tumefaciens A281 is encoded in the region of pTiBo542 outside the T-DNA. J Bacteriol 168:1291–1301
  • Hood EE, Gelvin SB, Melchers LS, Hoekema A (1993) New Agrobacterium helper plasmids for gene transfer to plants. Transgen Res 2:208–218
  • Hu Z, Wu YR, Li W, Gao HH (2006) Factors affecting Agrobacterium tumefaciens-mediated genetic transformation of Lycium barbarum L. In Vitro Cell Dev Biol Plant 42:461–466
  • Janssen BJ, Gardner RC (1993) The use of transient GUS expression to develop an Agrobacterium-mediated gene transfer system for kiwifruit. Plant Cell Rep 12:28–31
  • Jefferson RA, Kavanagh TA, Bevan NW (1987) GUS fusions: b-glucuronidase as a sensitive and versatile gene fusion marker in higher plants. EMBO J 6:3901–3907
  • Jime´nez VM (2001) Regulation of in vitro somatic embryogenesis with emphasis on the role of endogenous hormones. R Bras Fisiol Veg 13:196–223
  • Kaneda Y, Tabei S, Harada K, Akihama T, Kitamura K (1997) Combination of thidiazuron and basal media with low salt concentrations increases the frequency of shoot organogenesis in soybean [Glycine max (L.) Merr.]. Plant Cell Rep 17:8–12
  • Ko TS, Lee S, Krasnyanski S, Korban SS (2003) Two critical factors are required for efficient transformation of multiple soybean cultivars: Agrobacterium strain and orientation of immature cotyledon explant. Theor Appl Genet 107:439–447
  • Ko TS, Lee S, Farrand SK, Korban SS (2004a) A partially disarmed vir helper plasmid, pKYRT1, in conjunction with 2,4-dichlorophenoxyacetic acid promotes emergence of regenerable transgenic somatic embryos from immature cotyledons of soybean. Planta 218:536–541
  • Ko TS, Nelson RL, Korban SS (2004b) Screening multiple soybean cultivars (MG 00 to MG VIII) for somatic embryogenesis following Agrobacterium-mediated transformation of immature cotyledons. Crop Sci 44:1825–18314
  • Li DD, Shi W, Deng XX (2003) Factors influencing Agrobacteriummediated embryogenic callus transformation of Valencia sweet orange (Citrus sinensis) containing the pTA29-barnase gene. Tree Physiol 23:1209–1215
  • Liu HK, Yang C, Wei ZM (2004) Efficient Agrobacterium tumefaciens-mediated transformation of soybeans using an embryonic tip regeneration system. Planta 219:1042–1049
  • Loganathan M, Maruthasalam S, Shiu LY, Lien WC, Hsu WH, Lee PF, Yu CW, Lin CH (2010) Regeneration of soybean [Glycine max (L.) Merrill.] through direct somatic embryogenesis from the immature embryonic shoot tip. In Vitro Cell Dev Biol Plant 46:265–273
  • Mante S, Scorza R, Cordts J (1989) A simple, rapid protocol for adventitious shoot development from mature cotyledons of Glycine max cv. Bragg. In Vitro Cell Dev Biol Plant 25:385–388
  • Mante S, Morgens PH, Scorza R, Corts JM, Callahan AM (1991) Agrobacterium-mediated transformation of plum (Prunus domenstica L.) hypocotyls slices and regeneration of transgenic plant. Nat Biotechnol 9:853–857
  • Maughan PJ, Philip R, Cho MJ, Widholm JM, Vodkin LO (1999) Biolistic transformation, expression, and inheritance of bovine b-casein in soybean (Glycine max). In Vitro Cell Dev Biol Plant 35:334–349
  • McCabe DE, Swain WF, Martinell BJ, Christou P (1988) Stable transformation of soybean (Glycine max) by particle acceleration. Nat Biotechnol 6:923–926
  • McHughen A, Jordan M, Feist G (1989) A pre-culture period prior to Agrobacterium inoculation increases production of transgenic plants. J Plant Physiol 135:245–248
  • Meurer CA, Dinkins RD, Collins GB (1998) Factor affecting soybean cotyledonary node transformation. Plant Cell Rep 18:180–186
  • Murashige T, Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497
  • Olhoft PM, Somers DA (2001) L-Cysteine increases Agrobacteriummediated T-DNA delivery into soybean cotyledonary-node cells. Plant Cell Rep 20:706–711
  • Olhoft PM, Lin K, Galbraith J, Nielsen NC, Somers DA (2001) The role of thiol compounds in increasing Agrobacterium-mediated transformation of soybean cotyledonary-node cells. Plant Cell Rep 20:731–737
  • Parrot WA, Dryden G, Vogt S, Hildebrand DF, Collins GB, Williams EG (1988) Optimization of somatic embryogenesis and embryo germination in soybean. In Vitro Cell Dev Biol Plant 24:817–820
  • Parrot WA, Hoffman LM, Hildebrand DF, Williams EG, Collins GB (1989) Recovery of primary transformants of soybean. Plant Cell Rep 7:615–617
  • Paz MM, Martinez JC, Kalvig AB, Fonger TM, Wang K (2006) Improved cotyledonary node method using an alternative explant derived from mature seed for efficient Agrobacterium-mediated soybean transformation. Plant Cell Rep 25:206–213
  • Sairam RV, Franklin G, Hassel R, Smith B, Meeker K, Kashikar N, Parani M, Abed DA, Ismail S, Berry K, Goldman SL (2003) A study on the effect of genotypes, plant growth regulators and sugars in promoting plant regeneration via organogenesis from soybean cotyledonary nodal callus. Plant Cell Tissue Organ Cult 75:79–85
  • Samoylov VM, Tucker DM, Thibaud-Nissen F, Parrott WA (1998) A liquid-medium-based protocol for rapid regeneration from embryogenic soybean cultures. Plant Cell Rep 18:49–54
  • Sangwan RS, Bourgeois Y, Sangwan-Norreel BS (1991) Genetic transformation of Arabidopsis thaliana zygotic embryos and identification of critical parameters influencing transformation efficiency. Mol Gen Genet 230:475–485
  • Sangwan RS, Bourgeois Y, Brown S, Vasseur G, Sangwan-Noreel BS (1992) Characterization of competent cells and early events of Agrobacterium-mediated genetic transformation in Arabidopsis thaliana. Planta 188:439–456
  • Santare´m ER, Pelissier B, Finer JJ (1997) Effect of explant orientation, pH, solidifying agent and wounding on initiation of soybean somatic embryos. In Vitro Cell Dev Biol Plant 33:13–19
  • Santare´m ER, Trick HN, Essig JS, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean immature cotyledons: optimization of transient expression. Plant Cell Rep 17:752–759
  • Shetty K, Asano Y, Oosawa K (1992) Stimulation of in vitro shoot organogenesis in Glycine max (Merrill.) by allantoin and amides. Plant Sci 81:245–251
  • Shrawat AK, Becke D, Lorz H (2007) Agrobacterium tumefaciensmediated genetic transformation of barley (Hordeum vulgare L.). Plant Sci 172:281–290
  • Sreeramanan S, Maziah M, Abdullah MP, Sariah M, Xavier R (2006) Transient expression of gusA and gfp gene in Agrobacteriummediated banana transformation using single tiny meristematic bud. Asian J Plant Sci 5(3):468–480
  • Sriskandarajah S, Goodwin P (1998) Conditioning promotes regeneration and transformation in apple leaf explants. Plant Cell Tissue Organ Cult 53:1–11
  • Stewart CN Jr, Adang MJ, All JN, Boerma HR, Cardineau G, Tucker D, Parrot WA (1996) Genetic transformation, recovery, and characterization of fertile soybean transgenic for a synthetic Bacillus thuringiensis cryIAc gene. Plant Physiol 112:121–129
  • SubramaniamS, Samian R,Midrarullah, RathinamX (2009) Preliminary factors influencing transient expression of gusA in Dendrobium savin white protocorm-like bodies (PLBs) using Agrobacteriummediated transformation system.WASJ 7(10):1295–1307
  • Suzuki S, Nakano M (2002) Agrobacterium-mediated production of transgenic plants of Muscari armeniacum Leichtl.ex Bak. Plant Cell Rep 20:835–841
  • Tague B, Mantis J (2006) In-Planta Agrobacterium-mediated transformation by vacuum infiltration. Methods Mol Biol 323:215–223
  • Tomlin ES, Branch SR, Chamberlain D, Gabe H, Wright MS, Stewart CN Jr (2002) Screening of soybean [Glycine max (L.) Merrill.] lines for somatic embryo induction and maturation capability from immature cotyledons. In vitro Cell Dev Biol Plant 38:543–548
  • Torisky RS, Kovacs L, Avdiushko S, Newman JD, Hunt AG, Collins GB (1997) Development of a binary vector system for plant transformation based on the supervirulent Agrobacterium tumefaciens strain Chry5. Plant Cell Rep 17:102–108
  • Trick HN, Finer JJ (1997) SAAT: sonication-assisted Agrobacteriummediated transformation. Transgenic Res 6:329–337
  • Trick HN, Finer JJ (1998) Sonication-assisted Agrobacterium-mediated transformation of soybean [Glycine max (L.) Merrill.] embryogenic suspension culture tissue. Plant Cell Rep 17:482–488
  • Tsukazi H, Kuginuki Y, Aida R, Suzuki T (2002) Agrobacteriummediated transformation of a double haploid line of cabbage. Plant Cell Rep 21:257–262
  • Wang GL, Fang HJ (1998) Mechanism and technology of plant genetic engineering. Science Publisher, Beijing
  • Weir B, Gu X, Wang M, Upadhyaya N, Ellion A, Brettell RIS (2001) Agrobacterium tumefaciens-mediated transformation of wheat using suspension cells as a mode system and green fluorescent protein as a viral marker. Aust J Plant Physiol 28:307–318
  • Wright MS, Koehler SM, Hinchee MA, Carnes MG (1986) Plant regeneration by organogenesis in Glycine max. Plant Cell Rep 5:150–154
  • Yan B, Reddy MSS, Collins GB, Dinkins RD (2000) Agrobacterium tumefaciens-mediated transformation of soybean [Glycine max (L.) Merrill.] using immature zygotic cotyledon explants. Plant Cell Rep 19:1090–1097
  • Yang YS, Wada K, Futsuhara Y (1990) Comparative studies of organogenesis and plant regeneration in various soybean explants. Plant Sci 76:101–108
  • Yeh MS (1990) In vitro culture of immature soybean embryos II. The abilities of organogenesis and plantlet regeneration from different aged immature embryo in Glycine species. J Agric Assoc China 39:73–87
  • Yoshida T (2002) Adventitious shoot formation from hypocotyl sections of mature soybean seeds. Breed Sci 52:1–8
  • Zhang Z, Xing A, Staswick P, Clemente T (1999) The use of glufosinate as a selective agent in Agrobacteirum-mediated transformation of soybean. Plant Cell Tissue Organ Cult 56:37–46
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