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发根农杆菌pRi 1855 T-DNA介导的樱桃砧木Colt(甜樱桃×中国樱桃)转基因根的体细胞胚胎发生和芽再生

Somatic embryogenesis and shoot regeneration from transgenic roots of the cherry rootstock Colt (Prunus avium×P. pseudocerasus) mediated by pRi 1855 T-DNA of Agrobacterium rhizogenes.

作者信息

Gutièrrez-Pesce P, Taylor K, Muleo R, Rugini E

机构信息

Istituto di Ortofloroarboricoltura, Facoltà di Agraria, Università della Tuscia, Via S. Camillo De Lellis, I-01100 Viterbo, Italy, , , , , , IT.

Department of Plant Sciences, University of Arizona, 303 Forbes Building, Tucson, AZ 85721, USA, , , , , , US.

出版信息

Plant Cell Rep. 1998 Apr;17(6-7):574-580. doi: 10.1007/s002990050445.

DOI:10.1007/s002990050445
PMID:30736639
Abstract

Hairy roots were obtained after inoculation with Agrobacterium rhizogenes strain NCPPB 1855 of the in-vitro-grown shoots of the cherry rootstocks Colt (Prunus avium×P. pseudocerasus) and Mazzard F12/1 (P. avium L.). Not all putatively transgenic roots were able to grow in hormone-free medium. Mazzard F12/1 roots, induced with A. rhizogenes, did not differentiate any shoot or embryo, while both somatic embryos and shoots differentiated from the transgenic roots of Colt in medium containing 1 mg/l 6-benzylaminopurine and 1 mg/l 1-naphthaleneacetic acid. Somatic embryos were capable of secondary embryogenesis, but few developed into whole plants. DNA hybridization showed both a different number of bands and signal intensity in each of the five transgenic shoot clones and embryos examined. In a morphogenetic in vitro test, leaf explants of the transgenic shoot clones showed an increased capacity to differentiate roots, although clones differed in their sensitivity to the hormone ratio. Clones from the transgenic shoots had not only an increased rooting ability when grown in vitro but also exhibited various hairy root phenotypes when cultured in vitro and when transferred into the greenhouse.

摘要

用发根农杆菌菌株NCPPB 1855接种樱桃砧木科尔特(Prunus avium×P. pseudocerasus)和马扎德F12/1(P. avium L.)的离体培养芽后获得了毛状根。并非所有推定的转基因根都能在无激素培养基中生长。用发根农杆菌诱导的马扎德F12/1根没有分化出任何芽或胚,而在含有1 mg/l 6-苄基腺嘌呤和1 mg/l 1-萘乙酸的培养基中,体细胞胚和芽都从科尔特的转基因根中分化出来。体细胞胚能够进行次生胚发生,但很少发育成完整植株。DNA杂交显示,在所检测的五个转基因芽克隆和胚中,每个的条带数量和信号强度都不同。在形态发生的离体试验中,转基因芽克隆的叶片外植体表现出更强的生根分化能力,尽管不同克隆对激素比例的敏感性不同。转基因芽的克隆不仅在离体培养时生根能力增强,而且在离体培养和转移到温室中时还表现出各种毛状根表型。

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